Cat. No.: HY-14519

Methotrexate

(Synonyms: Amethopterin; CL14377; WR19039)

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Based on 77 publication(s) in Google Scholar

Methotrexate (Amethopterin), an antimetabolite and antifolate agent, inhibits the enzyme dihydrofolate reductase, thereby preventing the conversion of folic acid into tetrahydrofolate, and inhibiting DNA synthesis. Methotrexate, also an immunosuppressant and antineoplastic agent, is used for the research of rheumatoid arthritis and a number of different cancers (such as acute lymphoblastic leukemia).

For research use only. We do not sell to patients.

Purity: 99.95%
CAS No.: 59-05-2
Formula: C₂₀H₂₂N₈O₅
Molecular Weight:454.44

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4°C, protect from light

* In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

Methotrexate Chemical Structure

CAS No.: 59-05-2

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Biological Activity
- Description
- IC₅₀ & Target
- Cellular Effect
- In Vivo
- Clinical Trial
Chemical Information
Publications (77)
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Biological Activity

Description

IC₅₀ & Target

Traditional Cytotoxic Agents

All DNA/RNA Synthesis Isoforms :

All ADC Payloads Isoforms :

Cellular Effect

Cell Line	Type	Value	Description	References
143B	IC₅₀	8.8 nM Compound: MTX	Cytotoxic Activity was evaluated against 143B (TK-) tumor cells Cytotoxic Activity was evaluated against 143B (TK-) tumor cells	[PMID: 8632413]
5637	IC₅₀	0.016 μM Compound: Methotrexate	Cytotoxicity against human 5637 cells after 96 hrs by crystal violet staining Cytotoxicity against human 5637 cells after 96 hrs by crystal violet staining	[PMID: 19243173]
786-0	IC₅₀	33 nM Compound: MTX (1)	The IC50 value was measured on 786-0 cell line in renal tumor type. The IC50 value was measured on 786-0 cell line in renal tumor type.	[PMID: 9022795]
786-0	IC₅₀	33 nM Compound: MTX	Compound was evaluated for in vitro activity against 786-0 kidney cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against 786-0 kidney cell lines (Human tumor cells )	[PMID: 9857098]
A-375	IC₅₀	0.022 μM Compound: Methotrexate	Antiproliferative activity against human A375 cells Antiproliferative activity against human A375 cells	[PMID: 27217001]
A-427	IC₅₀	5.52 μM Compound: Methotrexate	Cytotoxicity against human A427 cells after 96 hrs by crystal violet staining Cytotoxicity against human A427 cells after 96 hrs by crystal violet staining	[PMID: 19243173]
A498	IC₅₀	0.022 μM Compound: Methotrexate	Antiproliferative activity against human A498 cells Antiproliferative activity against human A498 cells	[PMID: 27217001]
A498	IC₅₀	>1000 nM Compound: MTX	Compound was evaluated for in vitro activity against A498 kidney cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against A498 kidney cell lines (Human tumor cells )	[PMID: 9857098]
A549	IC₅₀	0.005 μg/mL Compound: ametropterin	Antitumor activity against human A549 cells Antitumor activity against human A549 cells	[PMID: 10579858]
A549	IC₅₀	0.02 μM Compound: 1a, MTX	Antiproliferative activity against A549 cells after 72 hrs in folate depleted media Antiproliferative activity against A549 cells after 72 hrs in folate depleted media	[PMID: 17127067]
A549	IC₅₀	0.1 μM Compound: 1a, MTX	Antiproliferative activity against A549 cells after 72 hrs by MTT assay Antiproliferative activity against A549 cells after 72 hrs by MTT assay	[PMID: 17127067]
A549	IC₅₀	35 ng/mL Compound: Methotrexate	In vitro growth inhibitory activity against A549 human nonsmall cell lung carcinoma cells. In vitro growth inhibitory activity against A549 human nonsmall cell lung carcinoma cells.	[PMID: 1847428]
A549	IC₅₀	37.4 nM Compound: 1, MTX	Antiproliferative activity against human A549 cells after 72 hrs by trypan blue exclusion method Antiproliferative activity against human A549 cells after 72 hrs by trypan blue exclusion method	[PMID: 20036565]
A549	IC₅₀	5 nM Compound: MTX	Antiproliferative activity against human A549 cells after 72 hrs by MTT assay Antiproliferative activity against human A549 cells after 72 hrs by MTT assay	[PMID: 20580561]
A549	IC₅₀	0.013 μM Compound: MTX	Cytotoxicity against human A549 cells assessed as growth inhibition after 72 hrs by MTT assay Cytotoxicity against human A549 cells assessed as growth inhibition after 72 hrs by MTT assay	[PMID: 25668494]
A549	IC₅₀	0.99 μM Compound: MTX	Antiproliferative activity against human A549 cells after 72 hrs by MTT assay in presence of leucovorin Antiproliferative activity against human A549 cells after 72 hrs by MTT assay in presence of leucovorin	[PMID: 27017552]
A549	IC₅₀	0.022 μM Compound: Methotrexate	Antiproliferative activity against human A549 cells Antiproliferative activity against human A549 cells	[PMID: 27217001]
A549	IC₅₀	0.25 μM Compound: MTX	Antiproliferative activity against human A549 cells after 72 hrs by MTT assay Antiproliferative activity against human A549 cells after 72 hrs by MTT assay	[PMID: 27886545]
A549	IC₅₀	0.0135 μM Compound: MTX	Antiproliferative activity against human A549 cells after 48 hrs by MTS assay Antiproliferative activity against human A549 cells after 48 hrs by MTS assay	[PMID: 28152430]
A549	GI₅₀	0.014 μM Compound: MTX	Antiproliferative activity against human A549 cells after 48 hrs by MTS assay Antiproliferative activity against human A549 cells after 48 hrs by MTS assay	[PMID: 28711701]
A549	IC₅₀	39.33 μg/mL Compound: MTX	Cytotoxicity against human A549 cells after 24 hrs by MTT assay Cytotoxicity against human A549 cells after 24 hrs by MTT assay	[PMID: 30554970]
A549	IC₅₀	990 nM Compound: MTX	Antiproliferative activity against human A549 cells assessed as reduction in cell viability after 72 hrs by MTT assay Antiproliferative activity against human A549 cells assessed as reduction in cell viability after 72 hrs by MTT assay	[PMID: 31200235]
A549	IC₅₀	0.014 μM Compound: MTX	Antiproliferative activity against human A549 cells assessed as reduction in cell viability by MTS assay Antiproliferative activity against human A549 cells assessed as reduction in cell viability by MTS assay	[PMID: 32058237]
A549	IC₅₀	31 nM Compound: MTX	Cytotoxic activity evaluated against A549 tumor cells Cytotoxic activity evaluated against A549 tumor cells	[PMID: 8632413]
A549	IC₅₀	23 nM Compound: MTX (1)	In vitro antitumor activity against A549 human non-small lung carcinoma cells containing 10% fetal bovine serum In vitro antitumor activity against A549 human non-small lung carcinoma cells containing 10% fetal bovine serum	[PMID: 9022795]
A549	IC₅₀	33 nM Compound: MTX	Compound was evaluated for in vitro activity against A549/ATCC lung cancer cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against A549/ATCC lung cancer cell lines (Human tumor cells )	[PMID: 9857098]
ACHN	IC₅₀	40 nM Compound: MTX (1)	The IC50 value was measured on ACHN cell line in renal tumor type. The IC50 value was measured on ACHN cell line in renal tumor type.	[PMID: 9022795]
ACHN	IC₅₀	40 nM Compound: MTX	Compound was evaluated for in vitro activity against ACHN kidney cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against ACHN kidney cell lines (Human tumor cells )	[PMID: 9857098]
Bel-7402	IC₅₀	82.3 μM Compound: MTX	Cytotoxicity against human Bel7402 cells after 48 hrs Cytotoxicity against human Bel7402 cells after 48 hrs	[PMID: 18555562]
BGC-823	IC₅₀	0.11 μM Compound: MTX	Cytotoxicity against human BGC823 cells after 48 hrs Cytotoxicity against human BGC823 cells after 48 hrs	[PMID: 18555562]
BT-549	IC₅₀	>1000 nM Compound: MTX	Compound was evaluated for in vitro activity against BT-549 breast cancer cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against BT-549 breast cancer cell lines (Human tumor cells )	[PMID: 9857098]
Caco-2	EC₅₀	1.1 μM Compound: Methotrexate	Cytotoxicity against human Caco-2 cells assessed as cell viability by XTT assay Cytotoxicity against human Caco-2 cells assessed as cell viability by XTT assay	[PMID: 20674353]
Caco-2	IC₅₀	0.32 μg/mL Compound: Methotrexate	Cytotoxicity against human Caco-2 cells after 48 hrs by MTT assay Cytotoxicity against human Caco-2 cells after 48 hrs by MTT assay	[PMID: 20846760]
Caco-2	IC₅₀	0.32 μg/mL Compound: Methotrexate	Cytotoxicity against human Caco2 cells after 48 hrs by MTT assay Cytotoxicity against human Caco2 cells after 48 hrs by MTT assay	10.1007/s00044-013-0773-3
CAKI-1	IC₅₀	>60 μg/mL Compound: methotrexate	Growth inhibition of human CAKI1 cells after 72 hrs Growth inhibition of human CAKI1 cells after 72 hrs	[PMID: 17569517]
CAKI-1	IC₅₀	25 μg/mL Compound: MTX	Inhibition of colony formation of Caki-1 cells by MTX Inhibition of colony formation of Caki-1 cells by MTX	[PMID: 2810330]
CAKI-1	IC₅₀	3 μg/mL Compound: MTX	Inhibition of colony formation of Caki-1 cells by hydrazone-linked conjugate (regiospecific coupling) of MTX and Dal K-20 Inhibition of colony formation of Caki-1 cells by hydrazone-linked conjugate (regiospecific coupling) of MTX and Dal K-20	[PMID: 2810330]
CAKI-1	IC₅₀	3.3 μg/mL Compound: MTX	Inhibition of colony formation of Caki-1 cells by amide-linked conjugate (regiospecific coupling) of MTX and Dal K-20 Inhibition of colony formation of Caki-1 cells by amide-linked conjugate (regiospecific coupling) of MTX and Dal K-20	[PMID: 2810330]
CAKI-1	IC₅₀	3.4 μg/mL Compound: MTX	Inhibition of colony formation of Caki-1 cells by hydrazone-linked conjugate of MTX and Dal K-20 Inhibition of colony formation of Caki-1 cells by hydrazone-linked conjugate of MTX and Dal K-20	[PMID: 2810330]
CAKI-1	IC₅₀	4.5 μg/mL Compound: MTX	Inhibition of colony formation of Caki-1 cells by amide-linked conjugate of MTX and Dal K-20 Inhibition of colony formation of Caki-1 cells by amide-linked conjugate of MTX and Dal K-20	[PMID: 2810330]
Cancer cell lines	IC₅₀	2.4 μM Compound: Methotrexate	Anticancer activity against human NCI lung cancer cell by MTT assay Anticancer activity against human NCI lung cancer cell by MTT assay	[PMID: 27112448]
Cancer cell lines	IC₅₀	2.2 μM Compound: Methotrexate	Anticancer activity against Homo sapiens (human) NCI lung cancer cell assessed as inhibition of cell proliferation after 24 hr by MTT assay Anticancer activity against Homo sapiens (human) NCI lung cancer cell assessed as inhibition of cell proliferation after 24 hr by MTT assay	10.1007/s00044-012-0260-2
carcinoma cell line	EC₅₀	13.8 nM Compound: MTX	Tested for their inhibition of growth of CCRF-CEM human leukemia on carcinoma cell lines. Tested for their inhibition of growth of CCRF-CEM human leukemia on carcinoma cell lines.	[PMID: 11052789]
carcinoma cell line	EC₅₀	14.5 nM Compound: MTX	Tested for their inhibition of growth of FaDu head on carcinoma cell lines. Tested for their inhibition of growth of FaDu head on carcinoma cell lines.	[PMID: 11052789]
CCRF S-180	IC₅₀	0.0054 μM Compound: MTX	Concentration required for 50% inhibition against S180 cells Concentration required for 50% inhibition against S180 cells	[PMID: 6403710]
CCRF S-180	ED₅₀	0.016 μM Compound: methotrexate	Cytotoxic activity against the S180 cell cultures Cytotoxic activity against the S180 cell cultures	[PMID: 6928967]
CCRF S-180	IC₅₀	10 nM Compound: MTX	Tested for the ability to inhibit growth in vitro against S180 cells Tested for the ability to inhibit growth in vitro against S180 cells	[PMID: 8277497]
CCRF-CEM	EC₅₀	14.4 nM Compound: MTX	Growth inhibition of CCRF-CEM human leukemia cells. Growth inhibition of CCRF-CEM human leukemia cells.	[PMID: 10956221]
CCRF-CEM	EC₅₀	13.8 nM Compound: MTX	Concentration required to inhibit growth against CCRF-CEM cell Concentration required to inhibit growth against CCRF-CEM cell	[PMID: 11052789]
CCRF-CEM	EC₅₀	15.5 nM Compound: MTX	Tested for their growth inhibition of CCRF-CEM human leukemia subline resistant as a result of decrease polyglutamylation. Tested for their growth inhibition of CCRF-CEM human leukemia subline resistant as a result of decrease polyglutamylation.	[PMID: 11052789]
CCRF-CEM	EC₅₀	2030 nM Compound: MTX	Tested for their growth inhibition of CCRF-CEM human leukemia subline resistant as a result of decrease in uptake. Tested for their growth inhibition of CCRF-CEM human leukemia subline resistant as a result of decrease in uptake.	[PMID: 11052789]
CCRF-CEM	EC₅₀	660 nM Compound: MTX	Tested for their growth inhibition of CCRF-CEM human leukemia subline resistant as a result of increase in wild type DHFR protein and activity. Tested for their growth inhibition of CCRF-CEM human leukemia subline resistant as a result of increase in wild type DHFR protein and activity.	[PMID: 11052789]
CCRF-CEM	EC₅₀	14 nM Compound: Methotrexate(MTX)	Growth inhibition of parental CCRF-CEM cells. Growth inhibition of parental CCRF-CEM cells.	[PMID: 11384244]
CCRF-CEM	EC₅₀	15 nM Compound: Methotrexate(MTX)	Growth inhibition of CCRF-CEM cells resistant to MTX Growth inhibition of CCRF-CEM cells resistant to MTX	[PMID: 11384244]
CCRF-CEM	EC₅₀	1900 nM Compound: Methotrexate(MTX)	Growth inhibition of DHFR-uptake, defined mechanisms of MTX resistance during continuous (0-120 h) exposure Growth inhibition of DHFR-uptake, defined mechanisms of MTX resistance during continuous (0-120 h) exposure	[PMID: 11384244]
CCRF-CEM	EC₅₀	600 nM Compound: Methotrexate(MTX)	Growth inhibition of DHFR-overexpressing cells, defined mechanisms of MTX resistance during continuous (0-120 h) exposure Growth inhibition of DHFR-overexpressing cells, defined mechanisms of MTX resistance during continuous (0-120 h) exposure	[PMID: 11384244]
CCRF-CEM	IC₅₀	0.004 μg/mL Compound: MTX	Compound was tested for cytotoxic activity in CCRF-CEM Human Leukemia cells by in vitro cytotoxicity assay Compound was tested for cytotoxic activity in CCRF-CEM Human Leukemia cells by in vitro cytotoxicity assay	[PMID: 11428931]
CCRF-CEM	EC₅₀	14.5 nM Compound: MTX	Inhibition of growth of CCRF-CEM human leukemia cells (n=4) Inhibition of growth of CCRF-CEM human leukemia cells (n=4)	[PMID: 11960504]
CCRF-CEM	EC₅₀	12 nM Compound: MTX	Concentration required to decrease cell growth by 50% in CCRF-CEM human leukemia cell line Concentration required to decrease cell growth by 50% in CCRF-CEM human leukemia cell line	[PMID: 12408727]
CCRF-CEM	EC₅₀	16 nM Compound: MTX	Concentration required to decrease cell growth by 50% in methotrexate-resistant CCRF-CEM human leukemia subline (R30dm) Concentration required to decrease cell growth by 50% in methotrexate-resistant CCRF-CEM human leukemia subline (R30dm)	[PMID: 12408727]
CCRF-CEM	EC₅₀	1700 nM Compound: MTX	Concentration required to decrease cell growth by 50% in methotrexate-resistant CCRF-CEM human leukemia subline (R2) Concentration required to decrease cell growth by 50% in methotrexate-resistant CCRF-CEM human leukemia subline (R2)	[PMID: 12408727]
CCRF-CEM	EC₅₀	595 nM Compound: MTX	Concentration required to decrease cell growth by 50% in methotrexate-resistant CCRF-CEM human leukemia subline (R1) Concentration required to decrease cell growth by 50% in methotrexate-resistant CCRF-CEM human leukemia subline (R1)	[PMID: 12408727]
CCRF-CEM	EC₅₀	13.7 nM Compound: Methotrexate	Growth inhibition of CCRF-CEM Human Leukemia Cell was determined Growth inhibition of CCRF-CEM Human Leukemia Cell was determined	[PMID: 12570380]
CCRF-CEM	EC₅₀	16.5 nM Compound: MTX	Growth inhibition of MTX-resistant human CCRF-CEM R30dm cells Growth inhibition of MTX-resistant human CCRF-CEM R30dm cells	[PMID: 15615522]
CCRF-CEM	EC₅₀	17.5 nM Compound: MTX	Growth inhibition of human CCRF-CEM cells Growth inhibition of human CCRF-CEM cells	[PMID: 15615522]
CCRF-CEM	EC₅₀	1500 nM Compound: MTX	Growth inhibition of methotrexate-transport-resistant human CCRF-CEM R2 cells after 120 hrs Growth inhibition of methotrexate-transport-resistant human CCRF-CEM R2 cells after 120 hrs	[PMID: 15615538]
CCRF-CEM	EC₅₀	660 nM Compound: MTX	Growth inhibition of methotrexate-resistant human CCRF-CEM R1 cells overproducing DHFR after 120 hrs Growth inhibition of methotrexate-resistant human CCRF-CEM R1 cells overproducing DHFR after 120 hrs	[PMID: 15615538]
CCRF-CEM	EC₅₀	7.9 nM Compound: MTX	Growth inhibition of methotrexate-resistant human CCRF-CEM R30dm cells expressing low levels of folyl-poly-glutamate synthetase after 120 hrs Growth inhibition of methotrexate-resistant human CCRF-CEM R30dm cells expressing low levels of folyl-poly-glutamate synthetase after 120 hrs	[PMID: 15615538]
CCRF-CEM	EC₅₀	8.2 nM Compound: MTX	Growth inhibition of human CCRF-CEM cells Growth inhibition of human CCRF-CEM cells	[PMID: 15615538]
CCRF-CEM	EC₅₀	13 nM Compound: Methotrexate	Growth inhibitory activity against CCRF-CEM human lymphoblastic leukemia cells Growth inhibitory activity against CCRF-CEM human lymphoblastic leukemia cells	[PMID: 16078850]
CCRF-CEM	EC₅₀	14.5 nM Compound: Methotrexate	Growth inhibitory activity against MTX resistant CCRF-CEM human lymphoblastic leukemia cells with decreased polyglutamylation Growth inhibitory activity against MTX resistant CCRF-CEM human lymphoblastic leukemia cells with decreased polyglutamylation	[PMID: 16078850]
CCRF-CEM	EC₅₀	1500 nM Compound: Methotrexate	Growth inhibitory activity against resistant CCRF-CEM human lymphoblastic leukemia cells with decreased uptake of MTX Growth inhibitory activity against resistant CCRF-CEM human lymphoblastic leukemia cells with decreased uptake of MTX	[PMID: 16078850]
CCRF-CEM	EC₅₀	620 nM Compound: Methotrexate	Growth inhibitory activity against MTX resistant subline of CCRF-CEM human lymphoblastic leukemia cells Growth inhibitory activity against MTX resistant subline of CCRF-CEM human lymphoblastic leukemia cells	[PMID: 16078850]
CCRF-CEM	EC₅₀	12.5 nM Compound: MTX	Growth inhibition of human CCRF-CEM cells Growth inhibition of human CCRF-CEM cells	[PMID: 16279780]
CCRF-CEM	EC₅₀	14.5 nM Compound: MTX	Growth inhibition of methotrexate-resistant human CCRF-CEM R30dm cells expressing low levels of folyl-poly-glutamate synthetase after 120 hrs Growth inhibition of methotrexate-resistant human CCRF-CEM R30dm cells expressing low levels of folyl-poly-glutamate synthetase after 120 hrs	[PMID: 16279780]
CCRF-CEM	EC₅₀	1500 nM Compound: MTX	Growth inhibition of methotrexate-transport-resistant human CCRF-CEM R2 cells after 120 hrs Growth inhibition of methotrexate-transport-resistant human CCRF-CEM R2 cells after 120 hrs	[PMID: 16279780]
CCRF-CEM	EC₅₀	615 nM Compound: MTX	Growth inhibition of methotrexate-resistant human CCRF-CEM R1 cells overproducing DHFR after 120 hrs Growth inhibition of methotrexate-resistant human CCRF-CEM R1 cells overproducing DHFR after 120 hrs	[PMID: 16279780]
CCRF-CEM	EC₅₀	14 nM Compound: MTX	Inhibition of growth in CCRF-CEM cells Inhibition of growth in CCRF-CEM cells	[PMID: 16451071]
CCRF-CEM	EC₅₀	17 nM Compound: MTX	Inhibition of growth in MTX-resistant CCRF-CEM cell line, R30dm cells Inhibition of growth in MTX-resistant CCRF-CEM cell line, R30dm cells	[PMID: 16451071]
CCRF-CEM	EC₅₀	13.3 nM Compound: Methotrexate, MTX	Growth inhibition of human CCRF-CEM cells Growth inhibition of human CCRF-CEM cells	[PMID: 17552508]
CCRF-CEM	EC₅₀	15.7 nM Compound: Methotrexate, MTX	Growth inhibition of of methotrexate-resistant human CCRF-CEM R30dm cells expressing low levels of FPGS Growth inhibition of of methotrexate-resistant human CCRF-CEM R30dm cells expressing low levels of FPGS	[PMID: 17552508]
CCRF-CEM	EC₅₀	1950 nM Compound: Methotrexate, MTX	Growth inhibition of human CCRF-CEM R2 cells with low methotrexate uptake Growth inhibition of human CCRF-CEM R2 cells with low methotrexate uptake	[PMID: 17552508]
CCRF-CEM	EC₅₀	410 nM Compound: Methotrexate, MTX	Growth inhibition of methotrexate-resistant human CCRF-CEM R1 cells overexpressing DHFR Growth inhibition of methotrexate-resistant human CCRF-CEM R1 cells overexpressing DHFR	[PMID: 17552508]
CCRF-CEM	EC₅₀	12.5 nM Compound: methotrexate, MTX	Cytotoxicity against human CCRF-CEM cells Cytotoxicity against human CCRF-CEM cells	[PMID: 18605720]
CCRF-CEM	IC₅₀	0.032 μM Compound: 1 (MTX)	Tested for cell-growth inhibition against human leukemic lymphoblast CEM cells Tested for cell-growth inhibition against human leukemic lymphoblast CEM cells	[PMID: 2898531]
CCRF-CEM	IC₅₀	6.6 μM Compound: 1 (MTX)	Tested for cell-growth inhibition against human leukemic lymphoblast CEM/MTX cells Tested for cell-growth inhibition against human leukemic lymphoblast CEM/MTX cells	[PMID: 2898531]
CCRF-CEM	IC₅₀	0.032 μM Compound: MTX	Cell growth inhibition against CEM cell from human leukemic lymphoblasts Cell growth inhibition against CEM cell from human leukemic lymphoblasts	[PMID: 3872941]
CCRF-CEM	IC₅₀	6.6 μM Compound: MTX	Cell growth inhibition against CEM/MTX cell from human leukemic lymphoblasts Cell growth inhibition against CEM/MTX cell from human leukemic lymphoblasts	[PMID: 3872941]
CCRF-CEM	IC₅₀	0.025 μM Compound: MTX	In vitro cytotoxicity by its growth inhibitory activity against human leukemic lymphoblasts (CEM cells). In vitro cytotoxicity by its growth inhibitory activity against human leukemic lymphoblasts (CEM cells).	[PMID: 6585550]
CCRF-CEM	EC₅₀	0.014 μM Compound: MTX	Growth inhibition against Human leukemic cell line(CCRF-CEM) Growth inhibition against Human leukemic cell line(CCRF-CEM)	[PMID: 7473577]
CCRF-CEM	EC₅₀	0.018 μM Compound: MTX	Growth inhibition against Human leukemic cell line (Methotrexate resistant CCRF-CEM) Growth inhibition against Human leukemic cell line (Methotrexate resistant CCRF-CEM)	[PMID: 7473577]
CCRF-CEM	EC₅₀	0.014 μM Compound: MTX	he compound was tested for Growth inhibition of the Human T-lymphoblastic Leukemia cell line CCRF-CEM he compound was tested for Growth inhibition of the Human T-lymphoblastic Leukemia cell line CCRF-CEM	[PMID: 7562910]
CCRF-CEM	EC₅₀	0.018 μM Compound: MTX	The compound was tested for Growth inhibition of Methotrexate-resistant human T-lymphoblastic leukemia cell line CCRF-CEM(sub line R30dm) The compound was tested for Growth inhibition of Methotrexate-resistant human T-lymphoblastic leukemia cell line CCRF-CEM(sub line R30dm)	[PMID: 7562910]
CCRF-CEM	IC₅₀	0.0007 μM Compound: MTX	Inhibitory concentration against CCRF-CEM leukemic cell DHFR(Dihydro folate reductase). Inhibitory concentration against CCRF-CEM leukemic cell DHFR(Dihydro folate reductase).	[PMID: 7562910]
CCRF-CEM	IC₅₀	0.7 nM Compound: MTX	Inhibitory concentration against CCRF-CEM leukemic cell DHFR(Dihydro folate reductase). Inhibitory concentration against CCRF-CEM leukemic cell DHFR(Dihydro folate reductase).	[PMID: 7562910]
CCRF-CEM	EC₅₀	14.5 nM Compound: MTX	Compound was evaluated for the growth inhibition of parental CCRF-CEM Compound was evaluated for the growth inhibition of parental CCRF-CEM	[PMID: 7783147]
CCRF-CEM	EC₅₀	14.5 nM Compound: MTX	Compound was evaluated for the growth inhibition of R30dm, a CCRF-CEM subline resistant to intermittent MTX exposure solely as a result of decreased polyglutamylation. Compound was evaluated for the growth inhibition of R30dm, a CCRF-CEM subline resistant to intermittent MTX exposure solely as a result of decreased polyglutamylation.	[PMID: 7783147]
CCRF-CEM	EC₅₀	3100 nM Compound: MTX	Compound was evaluated for the growth inhibition of R2, a CCRF-CEM subline resistant to MTX as a result of deffective uptake. Compound was evaluated for the growth inhibition of R2, a CCRF-CEM subline resistant to MTX as a result of deffective uptake.	[PMID: 7783147]
CCRF-CEM	EC₅₀	595 nM Compound: MTX	Compound was evaluated for the growth inhibition of R1, a CCRF-CEM subline resistant to MTX solely as a result of a 20-fold increase in wild-type DHFR protein and activity Compound was evaluated for the growth inhibition of R1, a CCRF-CEM subline resistant to MTX solely as a result of a 20-fold increase in wild-type DHFR protein and activity	[PMID: 7783147]
CCRF-CEM	IC₅₀	10.5 nM Compound: MTX	Tested for inhibitory concentration of cell growth against CCRF-CEM cell lines of human leukemic lymphoblast Tested for inhibitory concentration of cell growth against CCRF-CEM cell lines of human leukemic lymphoblast	[PMID: 8035423]
CCRF-CEM	IC₅₀	2550 nM Compound: MTX	Tested for inhibitory concentration of cell growth against CEM/MTX cell lines of human leukemic lymphoblast Tested for inhibitory concentration of cell growth against CEM/MTX cell lines of human leukemic lymphoblast	[PMID: 8035423]
CCRF-CEM	EC₅₀	16 nM Compound: MTX	the growth inhibition of, (during continuous exposure) human T-lymphoblastic leukemia cell line CCRF-CEM at resistance mechanism- sensitive the growth inhibition of, (during continuous exposure) human T-lymphoblastic leukemia cell line CCRF-CEM at resistance mechanism- sensitive	[PMID: 8164259]
CCRF-CEM	EC₅₀	2760 nM Compound: MTX	the growth inhibition of, (during continuous exposure) methotrexate resistant human T-lymphoblastic leukemia cell subline CEM/MTX at resistance mechanism - decreased influx the growth inhibition of, (during continuous exposure) methotrexate resistant human T-lymphoblastic leukemia cell subline CEM/MTX at resistance mechanism - decreased influx	[PMID: 8164259]
CCRF-CEM	ED₅₀	13.7 nM Compound: MTX	Compound was evaluated for growth inhibition of parental CCRF-CEM during continuous (120 h) exposure to MTX Compound was evaluated for growth inhibition of parental CCRF-CEM during continuous (120 h) exposure to MTX	[PMID: 8568828]
CCRF-CEM	ED₅₀	15.5 nM Compound: MTX	Growth inhibition of CCRF-CEM subline resistant to MTX solely as a result of decreased polyglutamylation during continuous (120 h) exposure. Growth inhibition of CCRF-CEM subline resistant to MTX solely as a result of decreased polyglutamylation during continuous (120 h) exposure.	[PMID: 8568828]
CCRF-CEM	ED₅₀	2550 nM Compound: MTX	Growth inhibition of CCRF-CEM subline resistant to MTX solely as a result of decreased MTX influx during continuous (120 h) exposure. Growth inhibition of CCRF-CEM subline resistant to MTX solely as a result of decreased MTX influx during continuous (120 h) exposure.	[PMID: 8568828]
CCRF-CEM	ED₅₀	655 nM Compound: MTX	Growth inhibition of CCRF-CEM subline resistant to MTX solely as a result of a 25-fold increase in wild-type DHFR activity during continuous (120 h) exposure to MTX Growth inhibition of CCRF-CEM subline resistant to MTX solely as a result of a 25-fold increase in wild-type DHFR activity during continuous (120 h) exposure to MTX	[PMID: 8568828]
CCRF-CEM	IC₅₀	0.72 μM Compound: MTX	Inhibitory activity against purified human dihydrofolate reductase (DHFR) in human leukemia cells (CCRF-CEM) Inhibitory activity against purified human dihydrofolate reductase (DHFR) in human leukemia cells (CCRF-CEM)	[PMID: 8568828]
CCRF-CEM	IC₅₀	14.5 μM Compound: MTX	Inhibitory activity against uptake of [3H]-MTX at Folyl-polyglutamate synthase by human leukemia cells (CCRF-CEM) Inhibitory activity against uptake of [3H]-MTX at Folyl-polyglutamate synthase by human leukemia cells (CCRF-CEM)	[PMID: 8568828]
CCRF-CEM	ED₅₀	14.5 nM Compound: MTX	Inhibitory activity against growth of CCRF-CEM human leukemia cells. Inhibitory activity against growth of CCRF-CEM human leukemia cells.	[PMID: 8691451]
CCRF-CEM	IC₅₀	0.82 nM Compound: MTX	Inhibitory concentration against dihydrofolate reductase (DHFR) enzyme isolated from CCRF-CEM human leukemia cells. Inhibitory concentration against dihydrofolate reductase (DHFR) enzyme isolated from CCRF-CEM human leukemia cells.	[PMID: 8691451]
CCRF-CEM	IC₅₀	3.2 μM Compound: MTX (Methotrexate)	Compound was evaluated for the inhibitory activity against Folyl-polyglutamate synthase from CCRF-CEM human leukemia cells. Compound was evaluated for the inhibitory activity against Folyl-polyglutamate synthase from CCRF-CEM human leukemia cells.	[PMID: 8863812]
CCRF-CEM	EC₅₀	14.3 nM Compound: MTX	Growth inhibition was measured as for inhibition of growth in CCRF-CEM cells. Growth inhibition was measured as for inhibition of growth in CCRF-CEM cells.	[PMID: 9554874]
CCRF-CEM	EC₅₀	567 nM Compound: MTX	Growth inhibition was measured for inhibition of growth in CCRF-CEM subline resistant to MTX solely as a result of a 20-fold increase in wild-type DHFR protein and activity. Growth inhibition was measured for inhibition of growth in CCRF-CEM subline resistant to MTX solely as a result of a 20-fold increase in wild-type DHFR protein and activity.	[PMID: 9554874]
CCRF-CEM	EC₅₀	8 nM Compound: MTX	Growth inhibition was measured for inhibition of growth in CCRF-CEM sublines resistant to MTX solely as a result of decreased polyglutamation. Growth inhibition was measured for inhibition of growth in CCRF-CEM sublines resistant to MTX solely as a result of decreased polyglutamation.	[PMID: 9554874]
CCRF-CEM	IC₅₀	29 nM Compound: MTX	Compound was evaluated for in vitro activity against CCRF-CEM leukemia cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against CCRF-CEM leukemia cell lines (Human tumor cells )	[PMID: 9857098]
COLO 205	IC₅₀	3.27 μM Compound: 6	Cytotoxicity against human COLO205 cells after 48 hrs by MTT assay Cytotoxicity against human COLO205 cells after 48 hrs by MTT assay	[PMID: 23968824]
COLO 205	IC₅₀	870 nM Compound: MTX	Compound was evaluated for in vitro activity against COLO 205 Colon cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against COLO 205 Colon cell lines (Human tumor cells )	[PMID: 9857098]
COLO 320DM	IC₅₀	5.25 μM Compound: 6	Cytotoxicity against human COLO320DM cells after 48 hrs by MTT assay Cytotoxicity against human COLO320DM cells after 48 hrs by MTT assay	[PMID: 23968824]
Colon 26	IC₅₀	>40 μM Compound: MTX	Inhibitory concentration of compound was tested against Colon 26 Mouse Colorectal Carcinoma on 4 hour exposure Inhibitory concentration of compound was tested against Colon 26 Mouse Colorectal Carcinoma on 4 hour exposure	[PMID: 8201595]
Colon 26	IC₅₀	31 nM Compound: MTX	Inhibitory concentration of compound was tested against Colon 26 tumor growth cell line Inhibitory concentration of compound was tested against Colon 26 tumor growth cell line	[PMID: 8201595]
DAN-G	IC₅₀	0.077 μM Compound: Methotrexate	Cytotoxicity against human DAN-G cells after 96 hrs by crystal violet staining Cytotoxicity against human DAN-G cells after 96 hrs by crystal violet staining	[PMID: 19243173]
Daoy	IC₅₀	9 nM Compound: MTX	Cytotoxic activity was evaluated against Daoy tumor cells Cytotoxic activity was evaluated against Daoy tumor cells	[PMID: 8632413]
DLD-1	IC₅₀	>60 μg/mL Compound: methotrexate	Growth inhibition of human DLD1 cells after 72 hrs Growth inhibition of human DLD1 cells after 72 hrs	[PMID: 17569517]
DU-145	IC₅₀	23 nM Compound: MTX (1)	The IC50 value was measured on DU-145 cell line in prostate tumor type. The IC50 value was measured on DU-145 cell line in prostate tumor type.	[PMID: 9022795]
DU-145	IC₅₀	45 nM Compound: MTX	Compound was evaluated for in vitro activity against DU-145 prostate cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against DU-145 prostate cell lines (Human tumor cells )	[PMID: 9857098]
Ehrlich	IC₅₀	0.0126 μM Compound: MTX	Concentration required for 50% inhibition against Ehrlich cells Concentration required for 50% inhibition against Ehrlich cells	[PMID: 6403710]
EKVX	IC₅₀	>1000 nM Compound: MTX (1)	The IC50 value was measured on EKVX cell line in NSCL tumor type. The IC50 value was measured on EKVX cell line in NSCL tumor type.	[PMID: 9022795]
EKVX	IC₅₀	>1000 nM Compound: MTX	Compound was evaluated for in vitro activity against EKVX lung cancer cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against EKVX lung cancer cell lines (Human tumor cells )	[PMID: 9857098]
EL4	IC₅₀	5.1 μM Compound: methotrexate	Cytotoxicity against mouse EL4 cells assessed as growth inhibition after 48 hrs by sulforhodamine B assay Cytotoxicity against mouse EL4 cells assessed as growth inhibition after 48 hrs by sulforhodamine B assay	[PMID: 25951057]
EL4	IC₅₀	1.57 μM Compound: MTX	Inhibitory growth of EL4 thymoma cell line from C57BL/6 mice Inhibitory growth of EL4 thymoma cell line from C57BL/6 mice	[PMID: 9379448]
Epidermoid carcinoma cell line	ED₅₀	0.001 μM Compound: MTX	Concentration required for inhibition of colony formation (cytotoxicity) against human epidermoid carcinoma cell number 2 Concentration required for inhibition of colony formation (cytotoxicity) against human epidermoid carcinoma cell number 2	[PMID: 6948961]
FaDu	EC₅₀	11.3 nM Compound: MTX	Growth inhibition of human squamous cell carcinoma FaDu cells during continuous Exposure (0-12 h) Growth inhibition of human squamous cell carcinoma FaDu cells during continuous Exposure (0-12 h)	[PMID: 10956221]
FaDu	EC₅₀	17 nM Compound: Methotrexate(MTX)	Growth inhibition of FaDu human head and neck squamous cell carcinoma cell lines. Growth inhibition of FaDu human head and neck squamous cell carcinoma cell lines.	[PMID: 11384244]
FaDu	EC₅₀	0.017 μM Compound: MTX	Growth inhibition against Human squamous cell line(Fadu) Growth inhibition against Human squamous cell line(Fadu)	[PMID: 7473577]
FaDu	EC₅₀	0.017 μM Compound: MTX	The compound was tested for Growth inhibition of the human squamous cell carcinoma FaDu cells The compound was tested for Growth inhibition of the human squamous cell carcinoma FaDu cells	[PMID: 7562910]
FaDu	EC₅₀	31 nM Compound: MTX	Growth inhibition of human squamous carcinoma cell lines following continuous (120 hours) exposure to MTX and compound FaDu cell line Growth inhibition of human squamous carcinoma cell lines following continuous (120 hours) exposure to MTX and compound FaDu cell line	[PMID: 7783147]
FaDu	EC₅₀	19 nM Compound: MTX	the growth inhibition of, (during continuous exposure) human squamous cell carcinoma FaDu at resistance mechanism - sensitive the growth inhibition of, (during continuous exposure) human squamous cell carcinoma FaDu at resistance mechanism - sensitive	[PMID: 8164259]
Fibroblast	IC₅₀	180 nM Compound: MTX, methotrexate	Cytotoxicity against mouse wild type fibroblast cells by MTT assay Cytotoxicity against mouse wild type fibroblast cells by MTT assay	[PMID: 17383876]
H35	IC₅₀	0.01 μM Compound: MTX	Growth inhibition of H35 cell lines Growth inhibition of H35 cell lines	[PMID: 6737432]
H35R0.3	IC₅₀	0.085 μM Compound: MTX	Compound was tested for growth inhibition of H35 Hepatoma cells. Compound was tested for growth inhibition of H35 Hepatoma cells.	[PMID: 1992121]
H35R0.3	IC₅₀	18 μM Compound: MTX	Compound was tested for inhibition of folinic acid transport in H35 Hepatoma cells. Compound was tested for inhibition of folinic acid transport in H35 Hepatoma cells.	[PMID: 1992121]
H35R0.3	IC₅₀	906 nM Compound: MTX	Inhibition of growth of H35R cells Inhibition of growth of H35R cells	[PMID: 2542557]
H35R0.3	IC₅₀	1.8 μM Compound: MTX	Growth inhibition of H35R0.3 cell lines Growth inhibition of H35R0.3 cell lines	[PMID: 6737432]
HBL-100	IC₅₀	0.04 μM Compound: MTX	Anticancer activity against Homo sapiens (human) HBL100 cells after 72 hr by MTT assay Anticancer activity against Homo sapiens (human) HBL100 cells after 72 hr by MTT assay	10.1007/s00044-012-0279-4
HCC 2998	IC₅₀	110 nM Compound: MTX	Compound was evaluated for in vitro activity against HCC2998 Colon cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against HCC2998 Colon cell lines (Human tumor cells )	[PMID: 9857098]
HCT-116	GI₅₀	>50 μM Compound: MTX	Growth inhibitory activity against human HCT116 cell line in presence of Hypoxanthine thymidine(HT) Growth inhibitory activity against human HCT116 cell line in presence of Hypoxanthine thymidine(HT)	[PMID: 15267250]
HCT-116	GI₅₀	0.01 μM Compound: MTX	Growth inhibitory activity against human HCT116 cell line Growth inhibitory activity against human HCT116 cell line	[PMID: 15267250]
HCT-116	GI₅₀	0.015 μM Compound: MTX	Cytotoxicity against human HCT116 cells after 72 hrs by MTT assay Cytotoxicity against human HCT116 cells after 72 hrs by MTT assay	[PMID: 26994844]
HCT-116	IC₅₀	0.75 μM Compound: MTX	Antiproliferative activity against human HCT116 cells after 72 hrs by MTT assay Antiproliferative activity against human HCT116 cells after 72 hrs by MTT assay	[PMID: 27886545]
HCT-116	GI₅₀	0.015 μM Compound: MTX	Antiproliferative activity against human HCT116 cells after 72 hrs by MTT assay Antiproliferative activity against human HCT116 cells after 72 hrs by MTT assay	[PMID: 28177228]
HCT-116	GI₅₀	0.13 μM Compound: MTX	Antiproliferative activity against human HCT116 cells after 48 hrs by MTS assay Antiproliferative activity against human HCT116 cells after 48 hrs by MTS assay	[PMID: 28711701]
HCT-116	IC₅₀	32.7 nM Compound: Methotrexate	Cytotoxicity against CD133 positive human HCT116 cells Cytotoxicity against CD133 positive human HCT116 cells	[PMID: 29468872]
HCT-116	IC₅₀	0.13 μM Compound: MTX	Antiproliferative activity against human HCT116 cells after 48 hrs by MTS assay Antiproliferative activity against human HCT116 cells after 48 hrs by MTS assay	[PMID: 29691154]
HCT-116	IC₅₀	9.25 μM Compound: MTX	Antiproliferative activity against human HCT-116 cells after 72 hrs by MTT assay Antiproliferative activity against human HCT-116 cells after 72 hrs by MTT assay	[PMID: 35964425]
HCT-116	IC₅₀	30 nM Compound: MTX (1)	The IC50 value was measured on HCT116 cell line in colon tumor type. The IC50 value was measured on HCT116 cell line in colon tumor type.	[PMID: 9022795]
HCT-116	IC₅₀	30 nM Compound: MTX	Compound was evaluated for in vitro activity against HCT116 Colon cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against HCT116 Colon cell lines (Human tumor cells )	[PMID: 9857098]
HCT-15	IC₅₀	30 nM Compound: MTX (1)	The IC50 value was measured on HCT-15 cell line in colon tumor type. The IC50 value was measured on HCT-15 cell line in colon tumor type.	[PMID: 9022795]
HCT-15	IC₅₀	30 nM Compound: MTX	Compound was evaluated for in vitro activity against HCT-15 Colon cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against HCT-15 Colon cell lines (Human tumor cells )	[PMID: 9857098]
HEK293	IC₅₀	0.022 μM Compound: Methotrexate	Cytotoxicity activity against HEK293 cells Cytotoxicity activity against HEK293 cells	[PMID: 27217001]
HEK293	IC₅₀	28.6 nM Compound: MTX	Cytotoxicity against human HEK293 cells assessed as reduction in cell viability incubated for 48 hrs in absence of ROS scavenger pyruvate by Alamar blue assay Cytotoxicity against human HEK293 cells assessed as reduction in cell viability incubated for 48 hrs in absence of ROS scavenger pyruvate by Alamar blue assay	[PMID: 31843461]
HEK293	IC₅₀	35 nM Compound: MTX	Cytotoxicity against human HEK293 cells assessed as reduction in cell viability incubated for 48 hrs in presence of ROS scavenger pyruvate by Alamar blue assay Cytotoxicity against human HEK293 cells assessed as reduction in cell viability incubated for 48 hrs in presence of ROS scavenger pyruvate by Alamar blue assay	[PMID: 31843461]
HEK293	IC₅₀	1.27 μM Compound: Methotrexate	Cytotoxicity against Homo sapiens (human) HEK293 cells assessed as inhibition of cell proliferation after 24 hr by MTT assay Cytotoxicity against Homo sapiens (human) HEK293 cells assessed as inhibition of cell proliferation after 24 hr by MTT assay	10.1007/s00044-012-0260-2
HeLa	IC₅₀	0.1 μM Compound: MTX	Cytotoxicity against human HeLa cells after 48 hrs Cytotoxicity against human HeLa cells after 48 hrs	[PMID: 18555562]
HeLa	IC₅₀	0.022 μM Compound: Methotrexate	Antiproliferative activity against human HeLa cells Antiproliferative activity against human HeLa cells	[PMID: 27217001]
HeLa	IC₅₀	>100 μM Compound: MTX	Antiproliferative activity against human HeLa cells after 48 hrs by MTS assay Antiproliferative activity against human HeLa cells after 48 hrs by MTS assay	[PMID: 28152430]
HeLa	GI₅₀	>100 μM Compound: MTX	Antiproliferative activity against human HeLa cells after 48 hrs by MTS assay Antiproliferative activity against human HeLa cells after 48 hrs by MTS assay	[PMID: 28711701]
HeLa	IC₅₀	>100 μM Compound: MTX	Antiproliferative activity against human HeLa cells after 48 hrs by MTS assay Antiproliferative activity against human HeLa cells after 48 hrs by MTS assay	[PMID: 29691154]
HeLa	IC₅₀	21 nM Compound: MTX	Cytotoxicity against human HeLa cells assessed as reduction in cell viability incubated for 48 hrs in absence of ROS scavenger pyruvate by Alamar blue assay Cytotoxicity against human HeLa cells assessed as reduction in cell viability incubated for 48 hrs in absence of ROS scavenger pyruvate by Alamar blue assay	[PMID: 31843461]
HeLa	IC₅₀	59.3 nM Compound: MTX	Cytotoxicity against human HeLa cells assessed as reduction in cell viability incubated for 48 hrs in presence of ROS scavenger pyruvate by Alamar blue assay Cytotoxicity against human HeLa cells assessed as reduction in cell viability incubated for 48 hrs in presence of ROS scavenger pyruvate by Alamar blue assay	[PMID: 31843461]
HeLa	IC₅₀	>100 μM Compound: MTX	Antiproliferative activity against human HeLa cells assessed as reduction in cell viability by MTS assay Antiproliferative activity against human HeLa cells assessed as reduction in cell viability by MTS assay	[PMID: 32058237]
HeLa	IC₅₀	27.94 μM Compound: Methotrexate	Antiproliferative activity against human HeLa cells assessed as reduction in cell viability after 24 hrs by MTT assay Antiproliferative activity against human HeLa cells assessed as reduction in cell viability after 24 hrs by MTT assay	[PMID: 32503691]
HeLa	IC₅₀	0.82 μM Compound: MTX	Antiproliferative activity against human HeLa cells assessed as reduction in cell viability preincubated for 12 hrs followed by 4 Gy irradiation and measured after 60 hrs by MTT assay Antiproliferative activity against human HeLa cells assessed as reduction in cell viability preincubated for 12 hrs followed by 4 Gy irradiation and measured after 60 hrs by MTT assay	[PMID: 32676149]
HeLa	IC₅₀	1.5 μM Compound: MTX	Antiproliferative activity against human HeLa cells assessed as reduction in cell viability incubated for 12 hrs by MTT assay Antiproliferative activity against human HeLa cells assessed as reduction in cell viability incubated for 12 hrs by MTT assay	[PMID: 32676149]
HeLa	IC₅₀	13.69 μM Compound: MTX	Antiproliferative activity against human HeLa cells after 72 hrs by MTT assay Antiproliferative activity against human HeLa cells after 72 hrs by MTT assay	[PMID: 35964425]
HEp-2	ED₅₀	0.0024 μM Compound: MTX	Effective dose against H.Ep.-2 cells Effective dose against H.Ep.-2 cells	[PMID: 7057425]
HEp-2	ED₅₀	2.4 nM Compound: methotrexate	Tested for cloning suppression test against H.Ep.-2 cells Tested for cloning suppression test against H.Ep.-2 cells	[PMID: 7365749]
HepG2	IC₅₀	0.99 μM Compound: MTX	Cytotoxicity against human HepG2 cells assessed as growth inhibition after 72 hrs by MTT assay Cytotoxicity against human HepG2 cells assessed as growth inhibition after 72 hrs by MTT assay	[PMID: 25668494]
HepG2	IC₅₀	15.8 μM Compound: methotrexate	Cytotoxicity against human HepG2 cells assessed as cell growth inhibition after 48 hrs by MTT assay Cytotoxicity against human HepG2 cells assessed as cell growth inhibition after 48 hrs by MTT assay	[PMID: 27186821]
HepG2	IC₅₀	0.022 μM Compound: Methotrexate	Antiproliferative activity against human HepG2 cells Antiproliferative activity against human HepG2 cells	[PMID: 27217001]
HepG2	IC₅₀	0.41 μM Compound: MTX	Antiproliferative activity against human HepG2 cells after 72 hrs by MTT assay Antiproliferative activity against human HepG2 cells after 72 hrs by MTT assay	[PMID: 27886545]
HepG2	IC₅₀	0.3 μM Compound: Methotrexate	Cytotoxicity against human HepG2 cells assessed as reduction in cell viability incubated for overnight by MTT assay Cytotoxicity against human HepG2 cells assessed as reduction in cell viability incubated for overnight by MTT assay	[PMID: 31260892]
HepG2	IC₅₀	21.9 μM Compound: MTX	Antiproliferative activity against human HepG2 cells after 72 hrs by MTT assay Antiproliferative activity against human HepG2 cells after 72 hrs by MTT assay	[PMID: 35964425]
HL-60	IC₅₀	0.012 μM Compound: methotrexate	Cytotoxicity against human HL60 cells after 72 hrs by MTT assay Cytotoxicity against human HL60 cells after 72 hrs by MTT assay	[PMID: 11170674]
HL-60	IC₅₀	6.92 nM Compound: methotrxate	Antiproliferative activity against human HL60 cells after 48 hrs by trypan blue exclusion test Antiproliferative activity against human HL60 cells after 48 hrs by trypan blue exclusion test	[PMID: 17497807]
HL-60	IC₅₀	0.21 μM Compound: MTX	Cytotoxicity against human HL60 cells after 48 hrs Cytotoxicity against human HL60 cells after 48 hrs	[PMID: 18555562]
HL-60	IC₅₀	1.09 μM Compound: MTX	Antiproliferative activity against human HL60 cells after 72 hrs by MTT assay Antiproliferative activity against human HL60 cells after 72 hrs by MTT assay	[PMID: 27886545]
HL-60	IC₅₀	0.0227 μM Compound: MTX	Antiproliferative activity against human HL60 cells after 48 hrs by MTS assay Antiproliferative activity against human HL60 cells after 48 hrs by MTS assay	[PMID: 28152430]
HL-60	GI₅₀	0.038 μM Compound: MTX	Antiproliferative activity against human HL60 cells after 48 hrs by MTS assay Antiproliferative activity against human HL60 cells after 48 hrs by MTS assay	[PMID: 28711701]
HL-60	IC₅₀	0.025 μM Compound: MTX	Antiproliferative activity against human HL60 cells after 48 hrs by MTS assay Antiproliferative activity against human HL60 cells after 48 hrs by MTS assay	[PMID: 29691154]
HL-60	IC₅₀	8.9 nM Compound: MTX	Cytotoxicity against human HL60 cells assessed as reduction in cell viability incubated for 48 hrs in absence of ROS scavenger pyruvate by Alamar blue assay Cytotoxicity against human HL60 cells assessed as reduction in cell viability incubated for 48 hrs in absence of ROS scavenger pyruvate by Alamar blue assay	[PMID: 31843461]
HL-60	IC₅₀	9.8 nM Compound: MTX	Cytotoxicity against human HL60 cells assessed reduction in cell viability incubated for 48 hrs in presence of ROS scavenger pyruvate by Alamar blue assay Cytotoxicity against human HL60 cells assessed reduction in cell viability incubated for 48 hrs in presence of ROS scavenger pyruvate by Alamar blue assay	[PMID: 31843461]
HL-60	IC₅₀	0.023 μM Compound: MTX	Antiproliferative activity against human HL-60 cells assessed as reduction in cell viability by MTS assay Antiproliferative activity against human HL-60 cells assessed as reduction in cell viability by MTS assay	[PMID: 32058237]
HL-60	ED₅₀	<0.05 μM Compound: MTX	Median-effective concentration for inhibition of human HL-60 cell growth at 48 h Median-effective concentration for inhibition of human HL-60 cell growth at 48 h	[PMID: 3373490]
HL-60	ED₅₀	<<0.05 μM Compound: MTX	Median-effective concentration for inhibition of human HL-60 cell growth at 72 h Median-effective concentration for inhibition of human HL-60 cell growth at 72 h	[PMID: 3373490]
HL-60	ED₅₀	0.1 μM Compound: MTX	Inhibition of [6-3H]-dUrd incorporation into DNA in HL-60 cells Inhibition of [6-3H]-dUrd incorporation into DNA in HL-60 cells	[PMID: 3373490]
HL-60	ED₅₀	609 μM Compound: MTX	Median-effective concentration for inhibition of human HL-60 cell growth at 24 h Median-effective concentration for inhibition of human HL-60 cell growth at 24 h	[PMID: 3373490]
HL-60	IC₅₀	8.1 nM Compound: MTX	Tested for the ability to inhibit growth in vitro against HL60 Tested for the ability to inhibit growth in vitro against HL60	[PMID: 8277497]
HL-60	IC₅₀	39 nM Compound: MTX	Compound was evaluated for in vitro activity against HL-60(TB) leukemia cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against HL-60(TB) leukemia cell lines (Human tumor cells )	[PMID: 9857098]
HL-60	IC₅₀	12 nM Compound: METHOTREXATE	Compound was tested for its anticancer activity against HL-60 cells. Compound was tested for its anticancer activity against HL-60 cells.	10.1016/S0960-894X(96)00486-6
HL-60	IC₅₀	0.012 μM Compound: Methotrexate (MTX)	Tested for the cytostatic activity as inhibitory concentration against leukemia HL-60 cells Tested for the cytostatic activity as inhibitory concentration against leukemia HL-60 cells	10.1016/S0960-894X(97)00071-1
HOP-92	IC₅₀	>1000 nM Compound: MTX	Compound was evaluated for in vitro activity against HOP-92 lung cancer cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against HOP-92 lung cancer cell lines (Human tumor cells )	[PMID: 9857098]
Hs-578T	IC₅₀	>1000 nM Compound: MTX	Compound was evaluated for in vitro activity against Hs 578.T breast cancer cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against Hs 578.T breast cancer cell lines (Human tumor cells )	[PMID: 9857098]
HSC-2	CC₅₀	>400 μM Compound: MTX	Cytotoxicity against human HSC-2 cells incubated for 48 hrs by MTT assay Cytotoxicity against human HSC-2 cells incubated for 48 hrs by MTT assay	[PMID: 33744685]
HT-29	IC₅₀	0.01 μg/mL Compound: ametropterin	Antitumor activity against human HT-29 cells Antitumor activity against human HT-29 cells	[PMID: 10579858]
HT-29	GI₅₀	>50 μM Compound: MTX	Growth inhibitory activity against human HT-29 cell line in presence of Hypoxanthine thymidine(HT) Growth inhibitory activity against human HT-29 cell line in presence of Hypoxanthine thymidine(HT)	[PMID: 15267250]
HT-29	GI₅₀	0.07 μM Compound: MTX	Growth inhibitory activity against human HT-29 cell line Growth inhibitory activity against human HT-29 cell line	[PMID: 15267250]
HT-29	IC₅₀	0.23 μg/mL Compound: Methotrexate	Cytotoxicity against human HT-29 cells after 48 hrs by MTT assay Cytotoxicity against human HT-29 cells after 48 hrs by MTT assay	[PMID: 20846760]
HT-29	GI₅₀	0.04 μM Compound: Methotrexate	Cytotoxicity against human HT-29 cells after 72 hrs by MTT assay Cytotoxicity against human HT-29 cells after 72 hrs by MTT assay	[PMID: 21115210]
HT-29	IC₅₀	1.4 μM Compound: MTX	Cytostatic activity against human HT-29 cells after 4 days by MTT assay Cytostatic activity against human HT-29 cells after 4 days by MTT assay	[PMID: 22480495]
HT-29	GI₅₀	0.04 μM Compound: Methotrexate	Cytotoxicity against human HT-29 cells after 72 hrs by MTT assay Cytotoxicity against human HT-29 cells after 72 hrs by MTT assay	[PMID: 23831811]
HT-29	IC₅₀	3.45 μM Compound: 6	Cytotoxicity against human HT-29 cells after 48 hrs by MTT assay Cytotoxicity against human HT-29 cells after 48 hrs by MTT assay	[PMID: 23968824]
HT-29	IC₅₀	4400 nM Compound: Methotrexate	Antiproliferative activity against human HT-29 cells after 96 hrs by MTT assay Antiproliferative activity against human HT-29 cells after 96 hrs by MTT assay	[PMID: 2778449]
HT-29	GI₅₀	>100 μM Compound: MTX	Antiproliferative activity against human HT-29 cells after 48 hrs by MTS assay Antiproliferative activity against human HT-29 cells after 48 hrs by MTS assay	[PMID: 28711701]
HT-29	IC₅₀	47.82 μg/mL Compound: MTX	Cytotoxicity against human HT-29 cells after 24 hrs by MTT assay Cytotoxicity against human HT-29 cells after 24 hrs by MTT assay	[PMID: 30554970]
HT-29	IC₅₀	>100 μM Compound: MTX	Antiproliferative activity against human HT29 cells assessed as reduction in cell viability by MTS assay Antiproliferative activity against human HT29 cells assessed as reduction in cell viability by MTS assay	[PMID: 32058237]
HT-29	IC₅₀	0.018 μM Compound: MTX	Inhibition of cell growth against HT-29 human gastrointestinal adenocarcinoma cell lines in vitro Inhibition of cell growth against HT-29 human gastrointestinal adenocarcinoma cell lines in vitro	[PMID: 3612694]
HT-29	IC₅₀	0.018 μM Compound: 1a	Evaluated for the inhibition of human gastrointestinal adenocarcinoma cells in vitro by HT-29 assay Evaluated for the inhibition of human gastrointestinal adenocarcinoma cells in vitro by HT-29 assay	[PMID: 3968685]
HT-29	IC₅₀	0.018 μM Compound: MTX	In vitro inhibition of growth of the human gastrointestinal adenocarcinoma cells HT-29 In vitro inhibition of growth of the human gastrointestinal adenocarcinoma cells HT-29	[PMID: 6694171]
HT-29	IC₅₀	32 nM Compound: MTX (1)	The IC50 value was measured on HT-29 cell line in colon tumor type. The IC50 value was measured on HT-29 cell line in colon tumor type.	[PMID: 9022795]
HT-29	IC₅₀	0.23 μg/mL Compound: Methotrexate	Cytotoxicity against human HT-29 cells after 48 hrs by MTT assay Cytotoxicity against human HT-29 cells after 48 hrs by MTT assay	10.1007/s00044-013-0773-3
HuTu80	IC₅₀	0.009 μM Compound: MTX	Inhibition of cell growth against HuTu 80 human gastrointestinal adenocarcinoma cell lines in vitro Inhibition of cell growth against HuTu 80 human gastrointestinal adenocarcinoma cell lines in vitro	[PMID: 3612694]
HuTu80	EC₅₀	4.5 nM Compound: MTX	Inhibition of growth (cytotoxicity) of HuTu 80 cell line in vitro Inhibition of growth (cytotoxicity) of HuTu 80 cell line in vitro	[PMID: 6410066]
HUVEC	IC₅₀	6.13 μM Compound: MTX	Cytotoxicity against HUVEC assessed as reduction in cell viability after 72 hrs by MTT assay Cytotoxicity against HUVEC assessed as reduction in cell viability after 72 hrs by MTT assay	[PMID: 31200235]
IGROV-1	IC₅₀	21 nM Compound: methotrexate	Antiproliferative activity against human RFC and FRalpha expressing human IGROV1 cells Antiproliferative activity against human RFC and FRalpha expressing human IGROV1 cells	[PMID: 18680275]
IGROV-1	IC₅₀	22 nM Compound: methotrexate	Antiproliferative activity against human RFC and FRalpha expressing human IGROV1 cells in presence of folic acid Antiproliferative activity against human RFC and FRalpha expressing human IGROV1 cells in presence of folic acid	[PMID: 18680275]
IGROV-1	IC₅₀	21 nM Compound: MTX	Antiproliferative activity against human IGROV1 cells expressing human RFC, FRalpha and PCFT assessed as reduction of viable cells after 96 hrs Antiproliferative activity against human IGROV1 cells expressing human RFC, FRalpha and PCFT assessed as reduction of viable cells after 96 hrs	[PMID: 21879757]
IGROV-1	IC₅₀	22 nM Compound: MTX	Antiproliferative activity against human IGROV1 cells expressing human RFC, FRalpha and PCFT assessed as reduction of viable cells after 96 hrs in the presence of 200 nM folic acid Antiproliferative activity against human IGROV1 cells expressing human RFC, FRalpha and PCFT assessed as reduction of viable cells after 96 hrs in the presence of 200 nM folic acid	[PMID: 21879757]
IGROV-1	IC₅₀	65 nM Compound: MTX	Compound was evaluated for in vitro activity against IGROV1 Ovary cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against IGROV1 Ovary cell lines (Human tumor cells )	[PMID: 9857098]
Jurkat	GI₅₀	1.25 μM Compound: Methotrexate	Growth inhibition of human Jurkat cells after 72 hrs by MTT assay Growth inhibition of human Jurkat cells after 72 hrs by MTT assay	[PMID: 21802949]
K562	IC₅₀	419 μM Compound: 1	In vitro cytotoxic concentration measured against human myelogenous leukemia K562 cells In vitro cytotoxic concentration measured against human myelogenous leukemia K562 cells	[PMID: 15780632]
K562	GI₅₀	0.03 μM Compound: Methotrexate	Cytotoxicity against human K562 cells after 72 hrs by MTT assay Cytotoxicity against human K562 cells after 72 hrs by MTT assay	[PMID: 21115210]
K562	IC₅₀	26 nM Compound: MTX	Compound was evaluated for in vitro activity against K562 leukemia cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against K562 leukemia cell lines (Human tumor cells )	[PMID: 9857098]
KB	IC₅₀	20 nM Compound: methotrexate	Antiproliferative activity against human RFC and FRalpha expressing human KB cells in presence of folic acid Antiproliferative activity against human RFC and FRalpha expressing human KB cells in presence of folic acid	[PMID: 18680275]
KB	IC₅₀	6 nM Compound: methotrexate	Antiproliferative activity against human RFC and FRalpha expressing human KB cells Antiproliferative activity against human RFC and FRalpha expressing human KB cells	[PMID: 18680275]
KB	IC₅₀	20 nM Compound: MTX	Antiproliferative activity against human KB cells expressing human RFC, FRalpha and PCFT assessed as reduction of viable cells after 96 hrs in the presence of 200 nM folic acid Antiproliferative activity against human KB cells expressing human RFC, FRalpha and PCFT assessed as reduction of viable cells after 96 hrs in the presence of 200 nM folic acid	[PMID: 21879757]
KB	IC₅₀	6 nM Compound: MTX	Antiproliferative activity against human KB cells expressing human RFC, FRalpha and PCFT assessed as reduction of viable cells after 96 hrs Antiproliferative activity against human KB cells expressing human RFC, FRalpha and PCFT assessed as reduction of viable cells after 96 hrs	[PMID: 21879757]
KB	IC₅₀	8.5 nM Compound: MTX	Inhibition of colony formation of human KB cells endogenously expressing RFC/FRalpha/PCFT incubated for 14 days by methylene blue staining based clonogenicty assay Inhibition of colony formation of human KB cells endogenously expressing RFC/FRalpha/PCFT incubated for 14 days by methylene blue staining based clonogenicty assay	[PMID: 22243528]
KB	IC₅₀	20.9 μM Compound: MTX	Antiproliferative activity against human KB cells after 48 hrs Antiproliferative activity against human KB cells after 48 hrs	[PMID: 23124219]
KB	IC₅₀	20 nM Compound: MTX	Growth inhibition of human KB cells expressing human RFC/FRalpha/PCFT after 96 hrs by CellTiter-blue assay in presence of folic acid Growth inhibition of human KB cells expressing human RFC/FRalpha/PCFT after 96 hrs by CellTiter-blue assay in presence of folic acid	[PMID: 24111942]
KB	IC₅₀	6 nM Compound: MTX	Growth inhibition of human KB cells expressing human RFC/FRalpha/PCFT after 96 hrs by CellTiter-blue assay Growth inhibition of human KB cells expressing human RFC/FRalpha/PCFT after 96 hrs by CellTiter-blue assay	[PMID: 24111942]
KB	IC₅₀	20 nM Compound: MTX	Cytotoxicity against human KB cells assessed as cell growth inhibition incubated up to 96 hrs in presence of 200 nM folic acid by Celltiter-blue cell viability assay Cytotoxicity against human KB cells assessed as cell growth inhibition incubated up to 96 hrs in presence of 200 nM folic acid by Celltiter-blue cell viability assay	[PMID: 25234128]
KB	IC₅₀	6 nM Compound: MTX	Cytotoxicity against human KB cells assessed as cell growth inhibition incubated up to 96 hrs by Celltiter-blue cell viability assay Cytotoxicity against human KB cells assessed as cell growth inhibition incubated up to 96 hrs by Celltiter-blue cell viability assay	[PMID: 25234128]
KB	IC₅₀	0.01 μM Compound: MTX	Cytotoxicity against human KB cells assessed as growth inhibition after 72 hrs by MTT assay Cytotoxicity against human KB cells assessed as growth inhibition after 72 hrs by MTT assay	[PMID: 25668494]
KB	IC₅₀	0.01 μM Compound: MTX	Antiproliferative activity against human KB cells after 72 hrs by MTT assay in presence of leucovorin Antiproliferative activity against human KB cells after 72 hrs by MTT assay in presence of leucovorin	[PMID: 27017552]
KB	IC₅₀	20 nM Compound: MTX	Antiproliferative activity against human KB cells expressing human RFC/FR-alpha/PCFT assessed as reduction in cell viability measured after 96 hrs in presence of folic acid by Cell-Titer Blue assay Antiproliferative activity against human KB cells expressing human RFC/FR-alpha/PCFT assessed as reduction in cell viability measured after 96 hrs in presence of folic acid by Cell-Titer Blue assay	[PMID: 29425443]
KB	IC₅₀	6 nM Compound: MTX	Antiproliferative activity against human KB cells expressing human RFC/FR-alpha/PCFT assessed as reduction in cell viability measured after 96 hrs by Cell-Titer Blue assay Antiproliferative activity against human KB cells expressing human RFC/FR-alpha/PCFT assessed as reduction in cell viability measured after 96 hrs by Cell-Titer Blue assay	[PMID: 29425443]
KB	IC₅₀	10 nM Compound: MTX	Antiproliferative activity against human KB cells assessed as reduction in cell viability after 72 hrs by MTT assay Antiproliferative activity against human KB cells assessed as reduction in cell viability after 72 hrs by MTT assay	[PMID: 31200235]
KB	IC₅₀	20 nM Compound: MTX	Inhibition of DHFR in human KB cells expressing RFC/FRalpha/PCFT assessed as reduction in cell growth after 96 hrs in presence of folic acid by Cell-Titer Blue assay Inhibition of DHFR in human KB cells expressing RFC/FRalpha/PCFT assessed as reduction in cell growth after 96 hrs in presence of folic acid by Cell-Titer Blue assay	[PMID: 32503687]
KB	IC₅₀	6 nM Compound: MTX	Inhibition of DHFR in human KB cells expressing RFC/FRalpha/PCFT assessed as reduction in cell growth after 96 hrs by Cell-Titer Blue assay Inhibition of DHFR in human KB cells expressing RFC/FRalpha/PCFT assessed as reduction in cell growth after 96 hrs by Cell-Titer Blue assay	[PMID: 32503687]
KB	ED₅₀	0.004 μg/mL Compound: 1, MTX	Cytotoxicity against human KB cells Cytotoxicity against human KB cells	[PMID: 448685]
KB	IC₅₀	19 nM Compound: MTX	Inhibitory concentration of compound was tested against KB tumor growth cell line Inhibitory concentration of compound was tested against KB tumor growth cell line	[PMID: 8201595]
KM12	IC₅₀	42 nM Compound: MTX (1)	The IC50 value was measured on KM-12 cell line in colon tumor type. The IC50 value was measured on KM-12 cell line in colon tumor type.	[PMID: 9022795]
KM12	IC₅₀	33 nM Compound: MTX	Compound was evaluated for in vitro activity against KM-12 Colon cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against KM-12 Colon cell lines (Human tumor cells )	[PMID: 9857098]
L1210	IC₅₀	0.0034 μM Compound: MTX	In vitro inhibition of L1210 cell growth In vitro inhibition of L1210 cell growth	[PMID: 1501226]
L1210	IC₅₀	3.9 nM Compound: Methotrexate	Concentration required to inhibit 50% growth of L1210 cells in culture. Concentration required to inhibit 50% growth of L1210 cells in culture.	[PMID: 1732549]
L1210	IC₅₀	4.51 nM Compound: MTX	Inhibitory concentration was evaluated as the concentration required for inhibition of L1210 cells. Inhibitory concentration was evaluated as the concentration required for inhibition of L1210 cells.	[PMID: 1732551]
L1210	IC₅₀	0.0046 μM Compound: 1	Ability to inhibit L1210 murine leukemia tumor cell growth in culture after 48 hr of its exposure. Ability to inhibit L1210 murine leukemia tumor cell growth in culture after 48 hr of its exposure.	[PMID: 1992118]
L1210	IC₅₀	200 μM Compound: 1	Ability to inhibit MTX-resistant L1210/R81 tumor cell growth was determined. Ability to inhibit MTX-resistant L1210/R81 tumor cell growth was determined.	[PMID: 1992118]
L1210	IC₅₀	0.009 μM Compound: Methotrexate	Compound was evaluated for cell growth inhibition of murine (L1210) leukemic cells Compound was evaluated for cell growth inhibition of murine (L1210) leukemic cells	[PMID: 1992122]
L1210	IC₅₀	4.6 nM Compound: methotrexate(MTX)	Compound was evaluated for the growth inhibition of L1210 cells Compound was evaluated for the growth inhibition of L1210 cells	[PMID: 1995880]
L1210	IC₅₀	5 nM Compound: Methotrexate (MTX)	Concentration inhibiting L1210 cell growth in culture Concentration inhibiting L1210 cell growth in culture	[PMID: 2296020]
L1210	IC₅₀	3.92 nM Compound: MTX	Inhibitory activity for the growth inhibition of L1210 cell line. Inhibitory activity for the growth inhibition of L1210 cell line.	[PMID: 2299633]
L1210	IC₅₀	2.55 nM Compound: MTX	Compound was evaluated for inhibitory effect on growth of L1210 cells at IC50 (n=4) Compound was evaluated for inhibitory effect on growth of L1210 cells at IC50 (n=4)	[PMID: 2423690]
L1210	IC₅₀	0.035 μM Compound: MTX	Compound was tested for its ability to inhibit dihydrofolate reductase purified from murine L1210 cells Compound was tested for its ability to inhibit dihydrofolate reductase purified from murine L1210 cells	[PMID: 2428979]
L1210	IC₅₀	0.2 μM Compound: MTX	Compound was tested for its ability to inhibit L1210 cell growth Compound was tested for its ability to inhibit L1210 cell growth	[PMID: 2428979]
L1210	IC₅₀	0.0044 μM Compound: MTX	Inhibition of the growth of methotrexate sensitive L1210 leukemia/S cells in culture Inhibition of the growth of methotrexate sensitive L1210 leukemia/S cells in culture	[PMID: 2704031]
L1210	IC₅₀	186 μM Compound: MTX	Inhibition of the growth of methotrexate resistant L1210 leukemia/R 81 cells in culture Inhibition of the growth of methotrexate resistant L1210 leukemia/R 81 cells in culture	[PMID: 2704031]
L1210	IC₅₀	0.002 μM Compound: MTX	Compound was tested for its ability to inhibit growth of L1210 mouse leukemia cells Compound was tested for its ability to inhibit growth of L1210 mouse leukemia cells	[PMID: 2871191]
L1210	IC₅₀	0.0046 μM Compound: 1 (MTX)	Tested for cell-growth inhibition against mouse leukemic L1210 cells Tested for cell-growth inhibition against mouse leukemic L1210 cells	[PMID: 2898531]
L1210	IC₅₀	0.025 μM Compound: 1 (MTX)	Tested for inhibition against purified Dihydrofolate reductase from L1210 murine leukemia cells Tested for inhibition against purified Dihydrofolate reductase from L1210 murine leukemia cells	[PMID: 2898531]
L1210	IC₅₀	0.009 μM Compound: 1 (MTX)	Inhibition of cell growth against L1210 murine leukemia cells Inhibition of cell growth against L1210 murine leukemia cells	[PMID: 2918496]
L1210	IC₅₀	6.0 x 10^-5 M Compound: MTX	Inhibition of thymidylate synthesis in permeabilised cells Inhibition of thymidylate synthesis in permeabilised cells	[PMID: 3091832]
L1210	IC₅₀	2.5 x 10^-9 M Compound: MTX	Compound was evaluated for growth inhibition in L1210 murine leukemia cells Compound was evaluated for growth inhibition in L1210 murine leukemia cells	[PMID: 3091834]
L1210	IC₅₀	0.005 nM Compound: Methotrexate	Inhibition of L1210 cultured murine leukemia cell growth Inhibition of L1210 cultured murine leukemia cell growth	[PMID: 3112397]
L1210	IC₅₀	200 nM Compound: Methotrexate	Inhibition of L1210/R81 murine leukemia cell growth Inhibition of L1210/R81 murine leukemia cell growth	[PMID: 3112397]
L1210	IC₅₀	3.9 x 10^-9 M Compound: MTX	Tested for growth inhibition of L1210 leukemia cells Tested for growth inhibition of L1210 leukemia cells	[PMID: 3121855]
L1210	IC₅₀	>2.0 x 10^-5 M Compound: MTX	Inhibition of GAR transformylase from mammalian L1210 cells Inhibition of GAR transformylase from mammalian L1210 cells	[PMID: 3184124]
L1210	IC₅₀	2.1 nM Compound: MTX	Evaluated for cell growth inhibition of L1210 cells in cultures Evaluated for cell growth inhibition of L1210 cells in cultures	[PMID: 3184124]
L1210	IC₅₀	20 μM Compound: MTX	Inhibitory activity against thymidylate synthase isolated from L1210 leukemia cells Inhibitory activity against thymidylate synthase isolated from L1210 leukemia cells	[PMID: 3339615]
L1210	IC₅₀	0.005 μM Compound: MTX	Inhibitory activity against cultured L1210 cells Inhibitory activity against cultured L1210 cells	[PMID: 3351853]
L1210	IC₅₀	0.0045 μM Compound: MTX	Inhibition of cell growth of murine L-1210 cells Inhibition of cell growth of murine L-1210 cells	[PMID: 3373490]
L1210	IC₅₀	0.0046 μM Compound: MTX	Inhibitory concentration against growth of L1210 cell line Inhibitory concentration against growth of L1210 cell line	[PMID: 3385730]
L1210	IC₅₀	0.002 μM Compound: MTX	Compound was tested for its inhibitory concentration to inhibit the growth of wild type L1210 cells. Compound was tested for its inhibitory concentration to inhibit the growth of wild type L1210 cells.	[PMID: 3462394]
L1210	IC₅₀	0.02 μM Compound: MTX	Compound was tested for its inhibitory concentration to inhibit the enzyme Dihydro Folate Reductase (DHFR) from murine L1210 leukemia cells. Compound was tested for its inhibitory concentration to inhibit the enzyme Dihydro Folate Reductase (DHFR) from murine L1210 leukemia cells.	[PMID: 3462394]
L1210	IC₅₀	0.03 μM Compound: MTX	Compound was tested for its inhibitory concentration to inhibit the growth of wild type L1210 cells; Range=0.01 to 0.03 Compound was tested for its inhibitory concentration to inhibit the growth of wild type L1210 cells; Range=0.01 to 0.03	[PMID: 3462394]
L1210	IC₅₀	220 μM Compound: MTX	Compound was tested for its inhibitory concentration to inhibit the growth of wild type L1210 cells and a subline (L1210 / R81). Compound was tested for its inhibitory concentration to inhibit the growth of wild type L1210 cells and a subline (L1210 / R81).	[PMID: 3462394]
L1210	IC₅₀	0.002 μM Compound: MTX	Cell growth inhibition against mouse leukemia L1210 cells Cell growth inhibition against mouse leukemia L1210 cells	[PMID: 3872941]
L1210	IC₅₀	19 μM Compound: MTX	Cell growth inhibition against mouse leukemia L1210/R71 cells Cell growth inhibition against mouse leukemia L1210/R71 cells	[PMID: 3872941]
L1210	IC₅₀	220 μM Compound: MTX	Cell growth inhibition against mouse leukemia L1210/R81 cells Cell growth inhibition against mouse leukemia L1210/R81 cells	[PMID: 3872941]
L1210	IC₅₀	0.012 μM Compound: 1a	Evaluated for the growth inhibition of L1210 /S cells Evaluated for the growth inhibition of L1210 /S cells	[PMID: 3968685]
L1210	IC₅₀	0.045 μM Compound: 1a	Evaluated for the inhibition of dihydrofolate reductase from L1210 mouse leukemia cells Evaluated for the inhibition of dihydrofolate reductase from L1210 mouse leukemia cells	[PMID: 3968685]
L1210	IC₅₀	1 μM Compound: 1a	Evaluated for the uptake of Methotrexate in L1210 leukemia cells Evaluated for the uptake of Methotrexate in L1210 leukemia cells	[PMID: 3968685]
L1210	IC₅₀	1.0 x 10^-8 M Compound: MTX	Inhibitory concentration against the growth of L1210 murine leukemia cells in tissue culture. Inhibitory concentration against the growth of L1210 murine leukemia cells in tissue culture.	[PMID: 4009615]
L1210	IC₅₀	3.28 nM Compound: MTX	Antitumor activity against L1210 cell in mice Antitumor activity against L1210 cell in mice	[PMID: 4020824]
L1210	IC₅₀	0.00084 μM Compound: MTX	Concentration required for 50% inhibition against L1210 cells Concentration required for 50% inhibition against L1210 cells	[PMID: 6403710]
L1210	IC₅₀	0.84 nM Compound: MTX	Concentration required for 50% inhibition against L1210 cells Concentration required for 50% inhibition against L1210 cells	[PMID: 6403710]
L1210	EC₅₀	3.2 nM Compound: MTX	Inhibition of growth (cytotoxicity) of L1210 cell line in vitro Inhibition of growth (cytotoxicity) of L1210 cell line in vitro	[PMID: 6410066]
L1210	IC₅₀	0.024 μM Compound: MTX	Growth inhibition of L1210 cell lines Growth inhibition of L1210 cell lines	[PMID: 6737432]
L1210	IC₅₀	50 nM Compound: MTX	Inhibitory activity towards dihydrofolate reductase in L1210 murine leukemia cell lines Inhibitory activity towards dihydrofolate reductase in L1210 murine leukemia cell lines	[PMID: 6737432]
L1210	IC₅₀	2.7 x 10^-9 M Compound: 4 (Methotrexate)	Inhibitory concentration of compound in growth of L-1210 cells Inhibitory concentration of compound in growth of L-1210 cells	[PMID: 6793726]
L1210	ED₅₀	0.016 μM Compound: methotrexate	Cytotoxic activity against the L1210 cell cultures Cytotoxic activity against the L1210 cell cultures	[PMID: 6928967]
L1210	IC₅₀	2.7 nM Compound: MTX	In vitro inhibition of L1210 cell growth in rodent neoplastic cells In vitro inhibition of L1210 cell growth in rodent neoplastic cells	[PMID: 7108907]
L1210	IC₅₀	9 nM Compound: MTX	Tested for the ability to inhibit growth in vitro against L1210 Tested for the ability to inhibit growth in vitro against L1210	[PMID: 8277497]
L1210	IC₅₀	9.5 nM Compound: MTX	Inhibition of growth in L1210 murine leukemia cells in culture expressed as IC50 (nM) Inhibition of growth in L1210 murine leukemia cells in culture expressed as IC50 (nM)	[PMID: 8340923]
L1210	IC₅₀	0.007 μM Compound: MTX (Methotrexate)	Compound was evaluated for the inhibition of L1210 murine leukemia cell growth.(In this study) Compound was evaluated for the inhibition of L1210 murine leukemia cell growth.(In this study)	[PMID: 8863812]
L1210	IC₅₀	0.073 μM Compound: MTX (Methotrexate)	compound was evaluated for the inhibitory activity against Dihydrofolate reductase in permeabilised L1210 cells. compound was evaluated for the inhibitory activity against Dihydrofolate reductase in permeabilised L1210 cells.	[PMID: 8863812]
L1210	IC₅₀	1.3 μM Compound: MTX (Methotrexate)	Compound was evaluated for the inhibition of L1210 murine leukemia cell growth.(From ref 12b) Compound was evaluated for the inhibition of L1210 murine leukemia cell growth.(From ref 12b)	[PMID: 8863812]
L1210	IC₅₀	20 nM Compound: MTX (Methotrexate)	Concentration required to inhibit the growth of L1210 cells Concentration required to inhibit the growth of L1210 cells	[PMID: 9022804]
L1210	IC₅₀	20 nM Compound: MTX	Concentration required to inhibit the growth of L1210 cells. Concentration required to inhibit the growth of L1210 cells.	[PMID: 9022805]
L1210 (R81)	IC₅₀	0.035 μM Compound: MTX	Compound was tested for its ability to inhibit purified dihydrofolate reductase(DHFR) from L1210/R81 cells Compound was tested for its ability to inhibit purified dihydrofolate reductase(DHFR) from L1210/R81 cells	[PMID: 2871191]
L1210 (R81)	IC₅₀	220 μM Compound: MTX	Compound was tested for its ability to inhibit growth of L1210/R81 cells Compound was tested for its ability to inhibit growth of L1210/R81 cells	[PMID: 2871191]
L1210 (R81)	IC₅₀	197 μM Compound: 1 (MTX)	Tested for cell-growth inhibition against mouse leukemic L1210/R81 cells Tested for cell-growth inhibition against mouse leukemic L1210/R81 cells	[PMID: 2898531]
L1210 (R81)	IC₅₀	25 nM Compound: Methotrexate	Inhibition of dihydrofolate reductase(DHFR) from L1210/R81 cells Inhibition of dihydrofolate reductase(DHFR) from L1210/R81 cells	[PMID: 3112397]
L1210 (R81)	IC₅₀	200 μM Compound: MTX	Inhibitory concentration against growth of L1210/R81 cell line Inhibitory concentration against growth of L1210/R81 cell line	[PMID: 3385730]
L1210 (R81)	IC₅₀	205 μM Compound: 1a	Evaluated for 50% inhibition of growth of L1210 / R81 cells (48 h), relative to untreated controls Evaluated for 50% inhibition of growth of L1210 / R81 cells (48 h), relative to untreated controls	[PMID: 3968685]
LCLC-103H cell line	IC₅₀	0.025 μM Compound: Methotrexate	Cytotoxicity against human LCLC-103H cells after 96 hrs by crystal violet staining Cytotoxicity against human LCLC-103H cells after 96 hrs by crystal violet staining	[PMID: 19243173]
LNCaP	IC₅₀	0.3 μM Compound: MTX	Cytostatic activity against human LNCAP cells after 4 days by MTT assay Cytostatic activity against human LNCAP cells after 4 days by MTT assay	[PMID: 22480495]
LOX IMVI	IC₅₀	0.057 μg/mL Compound: methotrexate	Growth inhibition of human LOX IMVI cells after 72 hrs Growth inhibition of human LOX IMVI cells after 72 hrs	[PMID: 17569517]
LOX IMVI	IC₅₀	26 nM Compound: MTX (1)	The IC50 value was measured on LOX IMVI cell line in melanoma tumor ty The IC50 value was measured on LOX IMVI cell line in melanoma tumor ty	[PMID: 9022795]
LOX IMVI	IC₅₀	26 nM Compound: MTX	Compound was evaluated for in vitro activity against LOXIMV1 Melanoma cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against LOXIMV1 Melanoma cell lines (Human tumor cells )	[PMID: 9857098]
M14	IC₅₀	32 nM Compound: MTX	Compound was evaluated for in vitro activity against M14 Melanoma cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against M14 Melanoma cell lines (Human tumor cells )	[PMID: 9857098]
M21	IC₅₀	108 μg/mL Compound: MTX	Concentration (20 ug/mL) of hydrazone-linked conjugate of MTX and Dal K-20 recquires to reduces the colony formation of melanoma M21 cells Concentration (20 ug/mL) of hydrazone-linked conjugate of MTX and Dal K-20 recquires to reduces the colony formation of melanoma M21 cells	[PMID: 2810330]
M21	IC₅₀	109 μg/mL Compound: MTX	Concentration (10 ug/mL) of amide-linked conjugate of MTX and Dal K-20 requires to reduces the colony formation of melanoma M21 cells Concentration (10 ug/mL) of amide-linked conjugate of MTX and Dal K-20 requires to reduces the colony formation of melanoma M21 cells	[PMID: 2810330]
M21	IC₅₀	109 μg/mL Compound: MTX	Concentration (15 ug/mL) of hydrazone-linked conjugate of MTX and Dal K-20 recquires to reduces the colony formation of melanoma M21 cells Concentration (15 ug/mL) of hydrazone-linked conjugate of MTX and Dal K-20 recquires to reduces the colony formation of melanoma M21 cells	[PMID: 2810330]
M21	IC₅₀	112 μg/mL Compound: MTX	Concentration (10 ug/mL) of hydrazone-linked conjugate of MTX and Dal K-20 recquires to reduces the colony formation of melanoma M21 cells Concentration (10 ug/mL) of hydrazone-linked conjugate of MTX and Dal K-20 recquires to reduces the colony formation of melanoma M21 cells	[PMID: 2810330]
M21	IC₅₀	112 μg/mL Compound: MTX	Concentration (20 ug/mL) of amide-linked conjugate (regiospecific coupling) of MTX and Dal K-20 required to reduce the colony formation of melanoma M21 cells Concentration (20 ug/mL) of amide-linked conjugate (regiospecific coupling) of MTX and Dal K-20 required to reduce the colony formation of melanoma M21 cells	[PMID: 2810330]
M21	IC₅₀	115 μg/mL Compound: MTX	Concentration (10 ug/mL) of amide-linked conjugate (regiospecific coupling) of MTX and Dal K-20 required to reduce the colony formation of melanoma M21 cells Concentration (10 ug/mL) of amide-linked conjugate (regiospecific coupling) of MTX and Dal K-20 required to reduce the colony formation of melanoma M21 cells	[PMID: 2810330]
M21	IC₅₀	115 μg/mL Compound: MTX	Concentration (20 ug/mL) of hydrazone-linked conjugate (regiospecific coupling) of MTX and Dal K-20 required to reduce the colony formation of melanoma M21 cells Concentration (20 ug/mL) of hydrazone-linked conjugate (regiospecific coupling) of MTX and Dal K-20 required to reduce the colony formation of melanoma M21 cells	[PMID: 2810330]
M21	IC₅₀	117 μg/mL Compound: MTX	Concentration (15 ug/mL) of amide-linked conjugate of MTX and Dal K-20 requires to reduces the colony formation of melanoma M21 cells Concentration (15 ug/mL) of amide-linked conjugate of MTX and Dal K-20 requires to reduces the colony formation of melanoma M21 cells	[PMID: 2810330]
M21	IC₅₀	118 μg/mL Compound: MTX	Concentration (15 ug/mL) of amide-linked conjugate (regiospecific coupling) of MTX and Dal K-20 required to reduce the colony formation of melanoma M21 cells Concentration (15 ug/mL) of amide-linked conjugate (regiospecific coupling) of MTX and Dal K-20 required to reduce the colony formation of melanoma M21 cells	[PMID: 2810330]
M21	IC₅₀	119 μg/mL Compound: MTX	Concentration (20 ug/mL) of amide-linked conjugate of MTX and Dal K-20 requires to reduces the colony formation of melanoma M21 cells Concentration (20 ug/mL) of amide-linked conjugate of MTX and Dal K-20 requires to reduces the colony formation of melanoma M21 cells	[PMID: 2810330]
M21	IC₅₀	120 μg/mL Compound: MTX	Concentration (10 ug/mL) of hydrazone-linked conjugate (regiospecific coupling) of MTX and Dal K-20 required to reduce the colony formation of melanoma M21 cells Concentration (10 ug/mL) of hydrazone-linked conjugate (regiospecific coupling) of MTX and Dal K-20 required to reduce the colony formation of melanoma M21 cells	[PMID: 2810330]
M21	IC₅₀	121 μg/mL Compound: MTX	Concentration (15 ug/mL) of hydrazone-linked conjugate (regiospecific coupling) of MTX and Dal K-20 required to reduce the colony formation of melanoma M21 cells Concentration (15 ug/mL) of hydrazone-linked conjugate (regiospecific coupling) of MTX and Dal K-20 required to reduce the colony formation of melanoma M21 cells	[PMID: 2810330]
Malme-3M	IC₅₀	>1000 nM Compound: MTX (1)	The IC50 value was measured on MALME-3M cell line in melanoma tumor type The IC50 value was measured on MALME-3M cell line in melanoma tumor type	[PMID: 9022795]
Malme-3M	IC₅₀	>1000 nM Compound: MTX	Compound was evaluated for in vitro activity against MALME-3M Melanoma cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against MALME-3M Melanoma cell lines (Human tumor cells )	[PMID: 9857098]
MC-38	IC₅₀	66 nM Compound: MTX	Inhibitory concentration of compound was tested against Colon 38 tumor growth cell line Inhibitory concentration of compound was tested against Colon 38 tumor growth cell line	[PMID: 8201595]
MCF7	IC₅₀	0.05 μM Compound: Methotrexate	Cytotoxicity against human MCF7 cells after 96 hrs by crystal violet staining Cytotoxicity against human MCF7 cells after 96 hrs by crystal violet staining	[PMID: 19243173]
MCF7	EC₅₀	0.9 μM Compound: Methotrexate	Cytotoxicity against human MCF7 cells assessed as cell viability by XTT assay Cytotoxicity against human MCF7 cells assessed as cell viability by XTT assay	[PMID: 20674353]
MCF7	GI₅₀	0.06 μM Compound: Methotrexate	Cytotoxicity against human MCF7 cells after 72 hrs by MTT assay Cytotoxicity against human MCF7 cells after 72 hrs by MTT assay	[PMID: 21115210]
MCF7	GI₅₀	0.06 μM Compound: Methotrexate	Cytotoxicity against human MCF7 cells after 72 hrs by MTT assay Cytotoxicity against human MCF7 cells after 72 hrs by MTT assay	[PMID: 23831811]
MCF7	GI₅₀	0.024 μM Compound: MTX	Cytotoxicity against human MCF7 cells after 72 hrs by MTT assay Cytotoxicity against human MCF7 cells after 72 hrs by MTT assay	[PMID: 26994844]
MCF7	IC₅₀	0.8 μM Compound: MTX; Methotrexate	Cytostatic activity against human MCF7 cells preincubated for 3 hrs followed by serum treatment for 72 hrs measured on day 4 by MTT assay Cytostatic activity against human MCF7 cells preincubated for 3 hrs followed by serum treatment for 72 hrs measured on day 4 by MTT assay	[PMID: 27031212]
MCF7	GI₅₀	0.024 μM Compound: MTX	Antiproliferative activity against human MCF7 cells after 72 hrs by MTT assay Antiproliferative activity against human MCF7 cells after 72 hrs by MTT assay	[PMID: 28177228]
MCF7	IC₅₀	0.097 μM Compound: MTX	Antiproliferative activity against human MCF7 cells in folate free medium after 72 hrs in presence of leucovorin by MTT assay Antiproliferative activity against human MCF7 cells in folate free medium after 72 hrs in presence of leucovorin by MTT assay	[PMID: 28830032]
MCF7	IC₅₀	31.48 μg/mL Compound: MTX	Cytotoxicity against human MCF7 cells after 24 hrs by MTT assay Cytotoxicity against human MCF7 cells after 24 hrs by MTT assay	[PMID: 30554970]
MCF7	IC₅₀	27.09 μM Compound: Methotrexate	Cytotoxicity against human MCF7 cells assessed as reduction in cell viability by MTT assay Cytotoxicity against human MCF7 cells assessed as reduction in cell viability by MTT assay	[PMID: 31404864]
MCF7	IC₅₀	49.22 μM Compound: Methotrexate	Antiproliferative activity against human MCF7 cells assessed as growth inhibition after 24 hrs by MTT assay Antiproliferative activity against human MCF7 cells assessed as growth inhibition after 24 hrs by MTT assay	[PMID: 32503691]
MCF7	IC₅₀	>22 μM Compound: MTX	Cytotoxicity against human MCF7 cells incubated for 72 hrs by SRB assay Cytotoxicity against human MCF7 cells incubated for 72 hrs by SRB assay	[PMID: 33479665]
MCF7	IC₅₀	0.015 μM Compound: MTX	Antineoplastic activity against human MCF7 cells incubated for 72 hrs by prestoblue reduction assay Antineoplastic activity against human MCF7 cells incubated for 72 hrs by prestoblue reduction assay	[PMID: 33479665]
MCF7	IC₅₀	25.32 μM Compound: MTX	Antiproliferative activity against human MCF7 cells after 72 hrs by MTT assay Antiproliferative activity against human MCF7 cells after 72 hrs by MTT assay	[PMID: 35964425]
MCF7	IC₅₀	36 nM Compound: MTX (1)	The IC50 value was measured on MCF-7 cell line in breast tumor type The IC50 value was measured on MCF-7 cell line in breast tumor type	[PMID: 9022795]
MCF7	IC₅₀	36 nM Compound: MTX	Compound was evaluated for in vitro activity against MCF-7 breast cancer cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against MCF-7 breast cancer cell lines (Human tumor cells )	[PMID: 9857098]
MCF7	IC₅₀	2.26 μM Compound: Methotrexate	Anticancer activity against Homo sapiens (human) MCF7 cells assessed as inhibition of cell proliferation after 24 hr by MTT assay Anticancer activity against Homo sapiens (human) MCF7 cells assessed as inhibition of cell proliferation after 24 hr by MTT assay	10.1007/s00044-012-0260-2
MDA-MB-231	EC₅₀	80 μM Compound: Methotrexate	Cytotoxicity against human MDA-MB-231 cells assessed as cell viability by XTT assay Cytotoxicity against human MDA-MB-231 cells assessed as cell viability by XTT assay	[PMID: 20674353]
MDA-MB-231	IC₅₀	0.5 μM Compound: 3; MTX	Cytotoxicity against human MDA-MB-231 cells expressing Y1R assessed as reduction in cell viability treated for 8 hrs followed by culturing in proliferation medium for 72 hrs by resazurin assay Cytotoxicity against human MDA-MB-231 cells expressing Y1R assessed as reduction in cell viability treated for 8 hrs followed by culturing in proliferation medium for 72 hrs by resazurin assay	[PMID: 26985967]
MDA-MB-231	IC₅₀	>100 μM Compound: MTX; Methotrexate	Cytostatic activity against human MDA-MB-231 cells preincubated for 3 hrs followed by serum treatment for 72 hrs measured on day 4 by MTT assay Cytostatic activity against human MDA-MB-231 cells preincubated for 3 hrs followed by serum treatment for 72 hrs measured on day 4 by MTT assay	[PMID: 27031212]
MDA-MB-231	IC₅₀	9.49 μM Compound: MTX	Antiproliferative activity against human MDA-MB-231 cells after 72 hrs by MTT assay Antiproliferative activity against human MDA-MB-231 cells after 72 hrs by MTT assay	[PMID: 27886545]
MDA-MB-435	IC₅₀	>1000 nM Compound: MTX	Compound was evaluated for in vitro activity against MDA-MB-435 breast cancer cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against MDA-MB-435 breast cancer cell lines (Human tumor cells )	[PMID: 9857098]
MDA-MB-468	IC₅₀	0.2 μM Compound: 3; MTX	Cytotoxicity against human MDA-MB-468 cells expressing Y1R assessed as reduction in cell viability treated for 8 hrs followed by culturing in proliferation medium for 72 hrs by resazurin assay Cytotoxicity against human MDA-MB-468 cells expressing Y1R assessed as reduction in cell viability treated for 8 hrs followed by culturing in proliferation medium for 72 hrs by resazurin assay	[PMID: 26985967]
MEF	IC₅₀	120 nM Compound: MTX, methotrexate	Cytotoxicity against FKBP deficient MEF cells by MTT assay Cytotoxicity against FKBP deficient MEF cells by MTT assay	[PMID: 17383876]
MOLT-4	IC₅₀	1.4 μM Compound: MTX	Compound was evaluated for inhibition of MTX influx into MOLT-4 cells Compound was evaluated for inhibition of MTX influx into MOLT-4 cells	[PMID: 1895294]
MOLT-4	IC₅₀	28 nM Compound: MTX	Compound was evaluated for in vitro activity against MOLT-4 leukemia cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against MOLT-4 leukemia cell lines (Human tumor cells )	[PMID: 9857098]
MOLT-4	IC₅₀	48 nM Compound: METHOTREXATE	Compound was tested for its anticancer activity against human T-lymphocytic leukemia MOLT-4 cells. Compound was tested for its anticancer activity against human T-lymphocytic leukemia MOLT-4 cells.	10.1016/S0960-894X(96)00486-6
NCI/ADR-RES	IC₅₀	78 nM Compound: MTX (1)	The IC50 value was measured on MCF7-ADR cell line in breast tumor type. The IC50 value was measured on MCF7-ADR cell line in breast tumor type.	[PMID: 9022795]
NCI/ADR-RES	IC₅₀	78 nM Compound: MTX	Compound was evaluated for in vitro activity against NCI/ADR-RES breast cancer cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against NCI/ADR-RES breast cancer cell lines (Human tumor cells )	[PMID: 9857098]
NCI-H1299	IC₅₀	0.072 μM Compound: MTX	Antiproliferative activity against human NCI-H1299 cells after 48 hrs by MTS assay Antiproliferative activity against human NCI-H1299 cells after 48 hrs by MTS assay	[PMID: 28152430]
NCI-H1299	GI₅₀	0.072 μM Compound: MTX	Antiproliferative activity against human NCI-H1299 cells after 48 hrs by MTS assay Antiproliferative activity against human NCI-H1299 cells after 48 hrs by MTS assay	[PMID: 28711701]
NCI-H1299	IC₅₀	0.072 μM Compound: MTX	Antiproliferative activity against human H1299 cells assessed as reduction in cell viability by MTS assay Antiproliferative activity against human H1299 cells assessed as reduction in cell viability by MTS assay	[PMID: 32058237]
NCI-H226	GI₅₀	10.25 μM Compound: Methotrexate	Growth inhibition of human NCI-H226 cells after 72 hrs by MTT assay Growth inhibition of human NCI-H226 cells after 72 hrs by MTT assay	[PMID: 21802949]
NCI-H226	IC₅₀	>1000 nM Compound: MTX	Compound was evaluated for in vitro activity against NCI-H226 lung cancer cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against NCI-H226 lung cancer cell lines (Human tumor cells )	[PMID: 9857098]
NCI-H23	IC₅₀	0.29 μg/mL Compound: methotrexate	Growth inhibition of human NCI-H23 cells after 72 hrs Growth inhibition of human NCI-H23 cells after 72 hrs	[PMID: 17569517]
NCI-H23	IC₅₀	43 nM Compound: MTX (1)	The IC50 value was measured on NCl-H23 cell line in NSCL tumor type. The IC50 value was measured on NCl-H23 cell line in NSCL tumor type.	[PMID: 9022795]
NCI-H23	IC₅₀	43 nM Compound: MTX	Compound was evaluated for in vitro activity against NCI-H23 lung cancer cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against NCI-H23 lung cancer cell lines (Human tumor cells )	[PMID: 9857098]
NCI-H460	IC₅₀	9.5 nM Compound: MTX	Cytotoxic Activity was evaluated against H460 tumor cells Cytotoxic Activity was evaluated against H460 tumor cells	[PMID: 8632413]
NCI-H460	IC₅₀	28 nM Compound: MTX (1)	The IC50 value was measured on NCl-H460 cell line in NSCL tumor type The IC50 value was measured on NCl-H460 cell line in NSCL tumor type	[PMID: 9022795]
NCI-H460	IC₅₀	28 nM Compound: MTX	Compound was evaluated for in vitro activity against NCI-H460 lung cancer cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against NCI-H460 lung cancer cell lines (Human tumor cells )	[PMID: 9857098]
NCI-H522	IC₅₀	229 nM Compound: MTX (1)	The IC50 value was measured on NCl-H522 cell line in NSCL tumor type. The IC50 value was measured on NCl-H522 cell line in NSCL tumor type.	[PMID: 9022795]
NCI-H522	IC₅₀	450 nM Compound: MTX	Compound was evaluated for in vitro activity against NCI-H522 lung cancer cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against NCI-H522 lung cancer cell lines (Human tumor cells )	[PMID: 9857098]
NIH3T3	IC₅₀	0.24 μg/mL Compound: Methotrexate	Cytotoxicity against mouse NIH/3T3 cells after 96 hrs by MTT assay Cytotoxicity against mouse NIH/3T3 cells after 96 hrs by MTT assay	10.1007/s00044-013-0773-3
OVCAR-3	IC₅₀	398 nM Compound: MTX (1)	The IC50 value was measured on OVCAR-3 cell line in ovarian tumor t The IC50 value was measured on OVCAR-3 cell line in ovarian tumor t	[PMID: 9022795]
OVCAR-3	IC₅₀	400 nM Compound: MTX	Compound was evaluated for in vitro activity against OVCAR-3 Ovary cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against OVCAR-3 Ovary cell lines (Human tumor cells )	[PMID: 9857098]
OVCAR-4	IC₅₀	>1000 nM Compound: MTX	Compound was evaluated for in vitro activity against OVCAR-4 Ovary cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against OVCAR-4 Ovary cell lines (Human tumor cells )	[PMID: 9857098]
OVCAR-5	IC₅₀	>1000 nM Compound: MTX (1)	The IC50 value was measured on OVCAR-5 cell line in ovarian tumor type. The IC50 value was measured on OVCAR-5 cell line in ovarian tumor type.	[PMID: 9022795]
OVCAR-5	IC₅₀	980 nM Compound: MTX	Compound was evaluated for in vitro activity against OVCAR-5 Ovary cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against OVCAR-5 Ovary cell lines (Human tumor cells )	[PMID: 9857098]
OVCAR-8	IC₅₀	31 nM Compound: MTX (1)	The IC50 value was measured on OVCAR-8 cell line in ovarian tumor type. The IC50 value was measured on OVCAR-8 cell line in ovarian tumor type.	[PMID: 9022795]
OVCAR-8	IC₅₀	31 nM Compound: MTX	Compound was evaluated for in vitro activity against OVCAR-8 Ovary cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against OVCAR-8 Ovary cell lines (Human tumor cells )	[PMID: 9857098]
P388	IC₅₀	0.002 μg/mL Compound: ametropterin	Antitumor activity against mouse P388 cells Antitumor activity against mouse P388 cells	[PMID: 10579858]
P388	IC₅₀	66 nM Compound: Methotrexate	Antiproliferative activity against mouse P388 cells after 48 hrs by MTT assay Antiproliferative activity against mouse P388 cells after 48 hrs by MTT assay	[PMID: 2778449]
P388	IC₅₀	223 nM Compound: MTX	Inhibitory concentration of compound was tested against P388:Methotrexate resistant cells tumor growth cell line Inhibitory concentration of compound was tested against P388:Methotrexate resistant cells tumor growth cell line	[PMID: 8201595]
P388	IC₅₀	23 nM Compound: MTX	Inhibitory concentration of compound was tested against P388 tumor growth cell line Inhibitory concentration of compound was tested against P388 tumor growth cell line	[PMID: 8201595]
P388	IC₅₀	6.6 nM Compound: MTX	Compound was tested for the inhibition of dihydrofolate reductase in P388 cells Compound was tested for the inhibition of dihydrofolate reductase in P388 cells	[PMID: 8201595]
P388	IC₅₀	4.8 nM Compound: MTX	Cytotoxic activity was evaluated against P388D1 tumor cells Cytotoxic activity was evaluated against P388D1 tumor cells	[PMID: 8632413]
PC-3	IC₅₀	0.027 μg/mL Compound: methotrexate	Growth inhibition of human PC3 cells after 72 hrs Growth inhibition of human PC3 cells after 72 hrs	[PMID: 17569517]
PC-3	IC₅₀	1 nM Compound: MTX	Antiproliferative activity against human PC3 cells after 72 hrs by MTT assay Antiproliferative activity against human PC3 cells after 72 hrs by MTT assay	[PMID: 20580561]
PC-3	GI₅₀	0.1 μM Compound: Methotrexate	Cytotoxicity against human PC3 cells after 72 hrs by MTT assay Cytotoxicity against human PC3 cells after 72 hrs by MTT assay	[PMID: 21115210]
PC-3	IC₅₀	28.78 μM Compound: MTX	Antiproliferative activity against human PC-3 cells after 72 hrs by MTT assay Antiproliferative activity against human PC-3 cells after 72 hrs by MTT assay	[PMID: 35964425]
PC-3	IC₅₀	2 nM Compound: MTX (1)	The IC50 value was measured on PC-3 cell line in prostate tumor type. The IC50 value was measured on PC-3 cell line in prostate tumor type.	[PMID: 9022795]
PC-3	IC₅₀	27 nM Compound: MTX	Compound was evaluated for in vitro activity against PC-3 prostate cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against PC-3 prostate cell lines (Human tumor cells )	[PMID: 9857098]
PPC-1	IC₅₀	0.01 μM Compound: 1a, MTX	Antiproliferative activity against PPC1 cells after 72 hrs in folate depleted media Antiproliferative activity against PPC1 cells after 72 hrs in folate depleted media	[PMID: 17127067]
PPC-1	IC₅₀	0.1 μM Compound: 1a, MTX	Antiproliferative activity against PPC1 cells after 72 hrs by MTT assay Antiproliferative activity against PPC1 cells after 72 hrs by MTT assay	[PMID: 17127067]
R1	EC₅₀	1050 nM Compound: MTX	Inhibition of growth of R1 (n=2) Inhibition of growth of R1 (n=2)	[PMID: 11960504]
R1	EC₅₀	720 nM Compound: MTX	the growth inhibition of, (during continuous exposure) methotrexate resistant human T-lymphoblastic leukemia cell subline R1 at resistance mechanism - increased DHFR the growth inhibition of, (during continuous exposure) methotrexate resistant human T-lymphoblastic leukemia cell subline R1 at resistance mechanism - increased DHFR	[PMID: 8164259]
R1-CCRF-CEM	EC₅₀	985 nM Compound: Methotrexate	Growth inhibition of MTX-resistant subline R1-CCRF-CEM Human Leukemia Cell was determined Growth inhibition of MTX-resistant subline R1-CCRF-CEM Human Leukemia Cell was determined	[PMID: 12570380]
R1-CCRF-CEM	EC₅₀	565 nM Compound: MTX	Inhibition of growth in MTX-resistant CCRF-CEM cell line, R1 cells Inhibition of growth in MTX-resistant CCRF-CEM cell line, R1 cells	[PMID: 16451071]
R2	EC₅₀	2600 nM Compound: MTX	Inhibition of growth of R2 (n=2) Inhibition of growth of R2 (n=2)	[PMID: 11960504]
R2	EC₅₀	2100 nM Compound: MTX	Growth inhibition of MTX-resistant human CCRF-CEM R2 cells Growth inhibition of MTX-resistant human CCRF-CEM R2 cells	[PMID: 15615522]
R2	IC₅₀	216 nM Compound: methotrexate	Antiproliferative activity against human RFC expressing Chinese hamster R2 cells Antiproliferative activity against human RFC expressing Chinese hamster R2 cells	[PMID: 18680275]
R2	IC₅₀	121 nM Compound: MTX	Antiproliferative activity against chinese hamster R2 cells expressing human PCFT assessed as reduction of viable cells after 96 hrs Antiproliferative activity against chinese hamster R2 cells expressing human PCFT assessed as reduction of viable cells after 96 hrs	[PMID: 21879757]
R2	IC₅₀	216 nM Compound: MTX	Antiproliferative activity against chinese hamster R2 cells assessed as reduction of viable cells after 96 hrs Antiproliferative activity against chinese hamster R2 cells assessed as reduction of viable cells after 96 hrs	[PMID: 21879757]
R2	IC₅₀	121 nM Compound: MTX	Growth inhibition of Chinese hamster R2 cells expressing human PCFT4 after 96 hrs by CellTiter-blue assay Growth inhibition of Chinese hamster R2 cells expressing human PCFT4 after 96 hrs by CellTiter-blue assay	[PMID: 24111942]
R2	IC₅₀	120.5 nM Compound: MTX	Cytotoxicity against chinese hamster R2 cells expressing human PCFT4 after 96 hrs by CellTitre-Blue fluorescence assay Cytotoxicity against chinese hamster R2 cells expressing human PCFT4 after 96 hrs by CellTitre-Blue fluorescence assay	[PMID: 24256410]
R2	IC₅₀	216 nM Compound: MTX	Cytotoxicity against Chinese hamster R2 cells assessed as cell growth inhibition incubated up to 96 hrs by Celltiter-blue cell viability assay Cytotoxicity against Chinese hamster R2 cells assessed as cell growth inhibition incubated up to 96 hrs by Celltiter-blue cell viability assay	[PMID: 25234128]
R2	IC₅₀	>1000 nM Compound: MTX	Cytotoxicity against Chinese hamster R2 cells expressing empty vector assessed as cell growth inhibition incubated up to 96 hrs by Celltiter-blue cell viability assay Cytotoxicity against Chinese hamster R2 cells expressing empty vector assessed as cell growth inhibition incubated up to 96 hrs by Celltiter-blue cell viability assay	[PMID: 25234128]
R2	IC₅₀	114 nM Compound: MTX	Cytotoxicity in RFC-null Chinese hamster R2 cells assessed as reduction in cell viability measured after 96 hrs by Cell-Titer Blue fluorescence analysis Cytotoxicity in RFC-null Chinese hamster R2 cells assessed as reduction in cell viability measured after 96 hrs by Cell-Titer Blue fluorescence analysis	[PMID: 27458733]
R2	IC₅₀	114 nM Compound: MTX	Cytotoxicity in RFC-null Chinese hamster R2 cells assessed as reduction in cell viability measured after 96 hrs by Cell-Titer Blue assay Cytotoxicity in RFC-null Chinese hamster R2 cells assessed as reduction in cell viability measured after 96 hrs by Cell-Titer Blue assay	[PMID: 29425443]
R2	IC₅₀	121 nM Compound: MTX	Inhibition of DHFR in Chinese Hamster MTXRII-OuaR2-4 R2 cells expressing human PCFT assessed as reduction in cell growth after 96 hrs by Cell-Titer Blue assay Inhibition of DHFR in Chinese Hamster MTXRII-OuaR2-4 R2 cells expressing human PCFT assessed as reduction in cell growth after 96 hrs by Cell-Titer Blue assay	[PMID: 32503687]
R2	IC₅₀	216 nM Compound: MTX	Inhibition of DHFR in Chinese Hamster MTXRII-OuaR2-4/FRalpha-null R2 cells assessed as reduction in cell growth after 96 hrs by Cell-Titer Blue assay Inhibition of DHFR in Chinese Hamster MTXRII-OuaR2-4/FRalpha-null R2 cells assessed as reduction in cell growth after 96 hrs by Cell-Titer Blue assay	[PMID: 32503687]
R2-CCRF-CEM	EC₅₀	2630 nM Compound: Methotrexate	Growth inhibition of MTX-resistant subline R2-CCRF-CEM Human Leukemia Cell was determined Growth inhibition of MTX-resistant subline R2-CCRF-CEM Human Leukemia Cell was determined	[PMID: 12570380]
R2-CCRF-CEM	EC₅₀	1700 nM Compound: MTX	Inhibition of growth in MTX-resistant CCRF-CEM cell line, R2 cells Inhibition of growth in MTX-resistant CCRF-CEM cell line, R2 cells	[PMID: 16451071]
R30dm-CCRF-CEM	EC₅₀	12.8 nM Compound: Methotrexate	Growth inhibition of MTX-resistant subline R30dm-CCRF-CEM Human Leukemia Cell was determined Growth inhibition of MTX-resistant subline R30dm-CCRF-CEM Human Leukemia Cell was determined	[PMID: 12570380]
R30dm-CCRF-CEM	EC₅₀	18 nM Compound: MTX	the growth inhibition of, (during continuous exposure) methotrexate resistant human T-lymphoblastic leukemia cell subline R30dm at resistance mechanism - decreased FPGS the growth inhibition of, (during continuous exposure) methotrexate resistant human T-lymphoblastic leukemia cell subline R30dm at resistance mechanism - decreased FPGS	[PMID: 8164259]
Raji	IC₅₀	0.04 μM Compound: methotrexate	Cytotoxicity against human Raji cells after 96 hrs by MTT assay Cytotoxicity against human Raji cells after 96 hrs by MTT assay	[PMID: 18513976]
RAW264.7	IC₅₀	159.2 μM Compound: Methotrexate	Cytotoxicity against mouse RAW264.7 cells assessed as inhibition of cell viability preincubated for 1 hr followed by LPS stimulation measured after 24 hrs by WST-1 assay Cytotoxicity against mouse RAW264.7 cells assessed as inhibition of cell viability preincubated for 1 hr followed by LPS stimulation measured after 24 hrs by WST-1 assay	[PMID: 31260892]
RAW264.7	IC₅₀	5.02 μM Compound: Methotrexate	Antiinflammatory activity against LPS-induced mouse RAW264.7 cells assessed as inhibition of NO overproduction preincubated for 1 hr prior to Escherichia coli LPS stimulation followed by further incubation for 24 hrs by Griess reagent based spectrophotome Antiinflammatory activity against LPS-induced mouse RAW264.7 cells assessed as inhibition of NO overproduction preincubated for 1 hr prior to Escherichia coli LPS stimulation followed by further incubation for 24 hrs by Griess reagent based spectrophotome	[PMID: 31260892]
RAW264.7	IC₅₀	1.23 μM Compound: Methotrexate	Antiproliferative activity against LPS-induced- mouse RAW264.7 cells assessed as inhibition of cell growth incubated for 24 hrs by CCK-8 assay Antiproliferative activity against LPS-induced- mouse RAW264.7 cells assessed as inhibition of cell growth incubated for 24 hrs by CCK-8 assay	[PMID: 38070351]
RPMI-8226	IC₅₀	33 nM Compound: MTX	Compound was evaluated for in vitro activity against RPMI-8226 leukemia cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against RPMI-8226 leukemia cell lines (Human tumor cells )	[PMID: 9857098]
RT-4	IC₅₀	0.04 μM Compound: Methotrexate	Cytotoxicity against human RT4 cells after 96 hrs by crystal violet staining Cytotoxicity against human RT4 cells after 96 hrs by crystal violet staining	[PMID: 19243173]
RXF 393	IC₅₀	>1000 nM Compound: MTX	Compound was evaluated for in vitro activity against RXF-393 kidney cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against RXF-393 kidney cell lines (Human tumor cells )	[PMID: 9857098]
SCC-15	IC₅₀	0.038 μM Compound: MTX	Inhibitory concentration against growth of SCC15 cell line Inhibitory concentration against growth of SCC15 cell line	[PMID: 3385730]
SCC-15	IC₅₀	0.58 μM Compound: MTX	Inhibitory concentration against growth of SCC15/R1 cell line Inhibitory concentration against growth of SCC15/R1 cell line	[PMID: 3385730]
SCC-25	IC₅₀	0.0075 μM Compound: MTX	Inhibitory concentration against growth of SCC25 cell line Inhibitory concentration against growth of SCC25 cell line	[PMID: 3385730]
SCC-25	IC₅₀	0.15 μM Compound: MTX	Inhibitory concentration against growth of SCC25/R1 cell line Inhibitory concentration against growth of SCC25/R1 cell line	[PMID: 3385730]
SCC-25	IC₅₀	7.5 nM Compound: MTX	Tested for inhibitory concentration against human SCC25 cell line Tested for inhibitory concentration against human SCC25 cell line	[PMID: 8035423]
SCC-25	IC₅₀	27 nM Compound: MTX (1)	In vitro antitumor activity against SCC25 human head and neck squamous carcinoma cells containing 10% fetal bovine serum In vitro antitumor activity against SCC25 human head and neck squamous carcinoma cells containing 10% fetal bovine serum	[PMID: 9022795]
SCC-7	IC₅₀	5.8 nM Compound: MTX	Tested for inhibitory concentration against SCC-VII murine squamous carcinoma cell line Tested for inhibitory concentration against SCC-VII murine squamous carcinoma cell line	[PMID: 8035423]
Sf21	IC₅₀	>1000 μM Compound: Methotrexate	Inhibition of human BSEP expressed in plasma membrane vesicles of Sf21 cells assessed as inhibition of ATP-dependent [3H]taurocholate uptake Inhibition of human BSEP expressed in plasma membrane vesicles of Sf21 cells assessed as inhibition of ATP-dependent [3H]taurocholate uptake	[PMID: 21965623]
Sf21	IC₅₀	>1000 μM Compound: Methotrexate	Inhibition of Sprague-Dawley rat Bsep expressed in plasma membrane vesicles of Sf21 cells assessed as inhibition of ATP-dependent [3H]taurocholate uptake Inhibition of Sprague-Dawley rat Bsep expressed in plasma membrane vesicles of Sf21 cells assessed as inhibition of ATP-dependent [3H]taurocholate uptake	[PMID: 21965623]
SF-268	IC₅₀	52 nM Compound: MTX (1)	The IC50 value was measured on SF-268 cell line in CNS tumor type. The IC50 value was measured on SF-268 cell line in CNS tumor type.	[PMID: 9022795]
SF-268	IC₅₀	52 nM Compound: MTX	Compound was evaluated for in vitro activity against SF-268 CNS cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against SF-268 CNS cell lines (Human tumor cells )	[PMID: 9857098]
SF-295	IC₅₀	36 nM Compound: MTX	Compound was evaluated for in vitro activity against SF-295 CNS cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against SF-295 CNS cell lines (Human tumor cells )	[PMID: 9857098]
SF-539	IC₅₀	35 nM Compound: MTX (1)	The IC50 value was measured on SF-539 cell line in CNS tumor type. The IC50 value was measured on SF-539 cell line in CNS tumor type.	[PMID: 9022795]
SF-539	IC₅₀	35 nM Compound: MTX	Compound was evaluated for in vitro activity against SF-539 CNS cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against SF-539 CNS cell lines (Human tumor cells )	[PMID: 9857098]
SK-MEL-2	IC₅₀	>1000 nM Compound: MTX	Compound was evaluated for in vitro activity against SK-MEL-2 Melanoma cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against SK-MEL-2 Melanoma cell lines (Human tumor cells )	[PMID: 9857098]
SK-MEL-28	IC₅₀	0.01 μg/mL Compound: ametropterin	Antitumor activity against human SK MEL28 cells Antitumor activity against human SK MEL28 cells	[PMID: 10579858]
SK-MEL-28	IC₅₀	>1000 nM Compound: MTX (1)	The IC50 value was measured on SKMEL-28 cell line in melanoma tumor type. The IC50 value was measured on SKMEL-28 cell line in melanoma tumor type.	[PMID: 9022795]
SK-MEL-28	IC₅₀	>1000 nM Compound: MTX	Compound was evaluated for in vitro activity against SK-MEL28 Melanoma cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against SK-MEL28 Melanoma cell lines (Human tumor cells )	[PMID: 9857098]
SK-MEL-5	IC₅₀	87 nM Compound: MTX (1)	The IC50 value was measured on SK-MEL-5 cell line in melanoma tumor type. The IC50 value was measured on SK-MEL-5 cell line in melanoma tumor type.	[PMID: 9022795]
SK-MEL-5	IC₅₀	87 nM Compound: MTX	Compound was evaluated for in vitro activity against SK-MEL-5 Melanoma cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against SK-MEL-5 Melanoma cell lines (Human tumor cells )	[PMID: 9857098]
SK-OV-3	IC₅₀	0.06 μM Compound: MTX	Antiproliferative activity against human SKOV3 cells after 48 hrs Antiproliferative activity against human SKOV3 cells after 48 hrs	[PMID: 23124219]
SNB-7	IC₅₀	>60 μg/mL Compound: methotrexate	Growth inhibition of human SNB7 cells after 72 hrs Growth inhibition of human SNB7 cells after 72 hrs	[PMID: 17569517]
SNB-75	IC₅₀	>1000 nM Compound: MTX (1)	The IC50 value was measured on SNB-75 cell line in CNS tumor type. The IC50 value was measured on SNB-75 cell line in CNS tumor type.	[PMID: 9022795]
SR	IC₅₀	33 nM Compound: MTX	Compound was evaluated for in vitro activity against SR leukemia cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against SR leukemia cell lines (Human tumor cells )	[PMID: 9857098]
SW 1116	IC₅₀	2.49 μM Compound: Methotrexate	Antitumor activity against human SW1116 cells assessed as reduction in cell viability incubated for overnight by MTT assay Antitumor activity against human SW1116 cells assessed as reduction in cell viability incubated for overnight by MTT assay	[PMID: 31260892]
SW480	IC₅₀	0.015 μM Compound: MTX	Inhibition of cell growth against SW480 human gastrointestinal adenocarcinoma cell lines in vitro Inhibition of cell growth against SW480 human gastrointestinal adenocarcinoma cell lines in vitro	[PMID: 3612694]
SW480	IC₅₀	0.015 μM Compound: 1a	Evaluated for the inhibition of human gastrointestinal adenocarcinoma cells in vitro by SW480 assay Evaluated for the inhibition of human gastrointestinal adenocarcinoma cells in vitro by SW480 assay	[PMID: 3968685]
SW480	IC₅₀	0.015 μM Compound: MTX	In vitro inhibition of growth of the human gastrointestinal adenocarcinoma cells SW480 In vitro inhibition of growth of the human gastrointestinal adenocarcinoma cells SW480	[PMID: 6694171]
SW-620	IC₅₀	0.013 μM Compound: MTX	Antiproliferative activity against human SW620 cells after 72 hrs by MTT assay in presence of leucovorin Antiproliferative activity against human SW620 cells after 72 hrs by MTT assay in presence of leucovorin	[PMID: 27017552]
SW-620	IC₅₀	0.022 μM Compound: MTX	Antiproliferative activity against human SW620 cells in folate free medium after 72 hrs in presence of leucovorin by MTT assay Antiproliferative activity against human SW620 cells in folate free medium after 72 hrs in presence of leucovorin by MTT assay	[PMID: 28830032]
SW-620	IC₅₀	13.1 nM Compound: MTX	Antiproliferative activity against human SW620 cells assessed as reduction in cell viability after 72 hrs by MTT assay Antiproliferative activity against human SW620 cells assessed as reduction in cell viability after 72 hrs by MTT assay	[PMID: 31200235]
SW-620	IC₅₀	33 nM Compound: MTX (1)	The IC50 value was measured on SW-620 cell line in colon tumor type. The IC50 value was measured on SW-620 cell line in colon tumor type.	[PMID: 9022795]
T47D	IC₅₀	0.16 μg/mL Compound: Methotrexate	Cytotoxicity against human T47D cells after 48 hrs by MTT assay Cytotoxicity against human T47D cells after 48 hrs by MTT assay	[PMID: 20846760]
T47D	IC₅₀	64.4 nM Compound: Methotrexate	Cytotoxicity against human T47D cells assessed as inhibition of cell viability incubated for 72 hrs by MTS assay Cytotoxicity against human T47D cells assessed as inhibition of cell viability incubated for 72 hrs by MTS assay	[PMID: 23468529]
T47D	IC₅₀	15 μM Compound: 3; MTX	Cytotoxicity against human T47D cells expressing Y1R assessed as reduction in cell viability treated for 8 hrs followed by culturing in proliferation medium for 72 hrs by resazurin assay Cytotoxicity against human T47D cells expressing Y1R assessed as reduction in cell viability treated for 8 hrs followed by culturing in proliferation medium for 72 hrs by resazurin assay	[PMID: 26985967]
T47D	IC₅₀	>1000 nM Compound: MTX	Compound was evaluated for in vitro activity against T-47D breast cancer cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against T-47D breast cancer cell lines (Human tumor cells )	[PMID: 9857098]
T47D	IC₅₀	0.16 μg/mL Compound: Methotrexate	Cytotoxicity against human T47D cells after 48 hrs by MTT assay Cytotoxicity against human T47D cells after 48 hrs by MTT assay	10.1007/s00044-013-0773-3
T-cell	IC₅₀	6.1 ng/mL Compound: MTX	Inhibitory activity against Con A-stimulated T cell proliferation. Inhibitory activity against Con A-stimulated T cell proliferation.	[PMID: 9379448]
TK-10	IC₅₀	>1000 nM Compound: MTX	Compound was evaluated for in vitro activity against TK-10 kidney cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against TK-10 kidney cell lines (Human tumor cells )	[PMID: 9857098]
U-251	IC₅₀	63 nM Compound: MTX	Compound was evaluated for in vitro activity against U251 CNS cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against U251 CNS cell lines (Human tumor cells )	[PMID: 9857098]
U-373MG ATCC	IC₅₀	12 nM Compound: MTX	Cytotoxic activity was evaluated against U373MG tumor cells Cytotoxic activity was evaluated against U373MG tumor cells	[PMID: 8632413]
U-87MG ATCC	IC₅₀	22 nM Compound: MTX	Cytotoxic activity was evaluated against U87MG tumor cells Cytotoxic activity was evaluated against U87MG tumor cells	[PMID: 8632413]
UACC-257	IC₅₀	790 nM Compound: MTX	Compound was evaluated for in vitro activity against UACC-257 Melanoma cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against UACC-257 Melanoma cell lines (Human tumor cells )	[PMID: 9857098]
UACC-62	IC₅₀	28 nM Compound: MTX (1)	The IC50 value was measured on UACC-62 cell line in melanoma tumor type. The IC50 value was measured on UACC-62 cell line in melanoma tumor type.	[PMID: 9022795]
UACC-62	IC₅₀	28 nM Compound: MTX	Compound was evaluated for in vitro activity against UACC-62 Melanoma cell lines (Human tumor cells ) Compound was evaluated for in vitro activity against UACC-62 Melanoma cell lines (Human tumor cells )	[PMID: 9857098]
UO-31	IC₅₀	191 nM Compound: MTX (1)	The IC50 value was measured on UO-31 cell line in renal tumor type. The IC50 value was measured on UO-31 cell line in renal tumor type.	[PMID: 9022795]
Vero	IC₅₀	9.2 nM Compound: MTX	Cytotoxic Activity was evaluated against Vero tumor cells Cytotoxic Activity was evaluated against Vero tumor cells	[PMID: 8632413]
W256	ED₅₀	0.02 μM Compound: methotrexate	Cytotoxic activity against the Walker 256 carcinosarcoma in rats Cytotoxic activity against the Walker 256 carcinosarcoma in rats	[PMID: 6928967]
WiDr	IC₅₀	0.025 μM Compound: MTX	Inhibition of cell growth against WIDR human gastrointestinal adenocarcinoma cell lines in vitro Inhibition of cell growth against WIDR human gastrointestinal adenocarcinoma cell lines in vitro	[PMID: 3612694]
WiDr	IC₅₀	0.025 μM Compound: 1a	Evaluated for the inhibition of human gastrointestinal adenocarcinoma cells in vitro by WIDR assay Evaluated for the inhibition of human gastrointestinal adenocarcinoma cells in vitro by WIDR assay	[PMID: 3968685]
WiDr	IC₅₀	0.025 μM Compound: MTX	In vitro inhibition of growth of the human gastrointestinal adenocarcinoma cells WIDR In vitro inhibition of growth of the human gastrointestinal adenocarcinoma cells WIDR	[PMID: 6694171]
WIL2	IC₅₀	0.0038 μM Compound: MTX	Inhibitory activity against dihydrofolate reductase (DHFR) obtained from human WIL2 cells Inhibitory activity against dihydrofolate reductase (DHFR) obtained from human WIL2 cells	[PMID: 3339615]
ZR-75-1	IC₅₀	>60 μg/mL Compound: methotrexate	Growth inhibition of human ZR-75-1 cells after 72 hrs Growth inhibition of human ZR-75-1 cells after 72 hrs	[PMID: 17569517]
ZR-75-1	IC₅₀	66 nM Compound: Methotrexate	Cytotoxicity against human ZR-75-1 cells assessed as inhibition of cell viability incubated for 72 hrs by MTS assay Cytotoxicity against human ZR-75-1 cells assessed as inhibition of cell viability incubated for 72 hrs by MTS assay	[PMID: 23468529]

In Vivo

Methotrexate (Amethopterin) reduces thymus and spleen indices of mice. Methotrexate markedly decreases white blood cells, thymic and splenic lymphocytes at dose ≥5 mg/kg. However, there is a significant difference between the treatment plus control group and the model group (p<0.01). The combination of grape seed proanthocyanidins and Siberian ginseng eleutherosides obviously diminishes the effects of Methotrexate exposure on indices of thymus and spleens in mice^[2].
Methotrexate (MTX) (2 mg/kg; i.p.; once in a week for 5 weeks) is effective in Freund's complete adjuvant-induced arthritis. The combination of Methotrexate (1 mg/kg; i.p.; once in a week for 5 weeks) and Curcumin (30 mg/kg and 100 mg/kg, thrice a week for 5 weeks; i.p.) shows a significant anti-arthritic action and protection from hematological toxicity^[4].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Clinical Trial

NCT Number	Sponsor	Condition	Start Date	Phase A549
NCT01329991	Plexxikon\|		2011-05	PHASE1

Chemical Information

CAS No. 59-05-2
Appearance Solid
Molecular Weight 454.44
Formula C₂₀H₂₂N₈O₅
Color Light yellow to yellow
SMILES

NC1=NC(N)=C2C(N=CC(CN(C)C3=CC=C(C(N[C@@H](CCC(O)=O)C(O)=O)=O)C=C3)=N2)=N1
Synonyms

Amethopterin; CL14377; WR19039
Shipping

Room temperature in continental US; may vary elsewhere.
Storage

4°C, protect from light

* In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

Publications (77)

All Cell Imaging/StainingCell Proliferation/Viability AssayRT-PCRCell Migration/Invasion Assay

Sort by:

Journal Impact Factor

Mol Cancer

Deubiquitination of CDC6 by OTUD6A promotes tumour progression and chemoresistance. [Abstract]

2024 Apr 29;23(1):86. PMID: 38685067
Mol Cancer

Organoids derived from patients provide a new opportunity for research and individualized treatment of malignant peritoneal mesothelioma. [Abstract]

2024 Jan 10;23(1):12. PMID: 38200517
Adv Mater

3D Bioprinted Human Synovium-Cartilage Models Mimic Rheumatoid Arthritis Microenvironment and Recapitulate In Vivo Therapeutic Responses. [Abstract]

2025 Dec 12:e13952. PMID: 41388589
Adv Mater

Soft Extrudable Dendritic Particles with Nanostructured Tendrils for Local Adhesion and Drug Release to Bladder Cancers. [Abstract]

2025 Jul 4:e2505231. PMID: 40611758
Nat Commun

ACSS2 drives senescence-associated secretory phenotype by limiting purine biosynthesis through PAICS acetylation. [Abstract]

2025 Feb 28;16(1):2071. PMID: 40021646

Methotrexate purchased from MedChemExpress. Usage Cited in: Nat Commun. 2025 Feb 28;16(1):2071. [Abstract]

OIS cells were infected with shCtrl orshACSS2 followed by selection and treated with or without MTX (40 μM) for 2 days. Cells were stained with γH2AX.

Methotrexate purchased from MedChemExpress. Usage Cited in: Nat Commun. 2025 Feb 28;16(1):2071. [Abstract]

OIS cells were infected with shCtrl orshACSS2 followed by selection and treated with or without MTX (40 μM) for 2 days. The expression of SASP genes was analysed by qRT–PCR.
J Clin Invest

Therapeutic targeting of metabolic vulnerabilities in cancers with MLL3/4-COMPASS epigenetic regulator mutations. [Abstract]

2023 Jul 3;133(13):e169993. PMID: 37252797

Methotrexate purchased from MedChemExpress. Usage Cited in: J Clin Invest. 2023 Jul 3;133(13):e169993. [Abstract]

A CellTiter-Glo® luminescent cell viability assay was performed on CAL51 WT and MLL4 KO cells, which were treated with 0, 0.04, or 0.4 μM Methotrexate (MTX) in the presence of H2O, 50 μM thymidine, or 50 μM inosine.
J Adv Res

Columbianadin ameliorates rheumatoid arthritis by attenuating synoviocyte hyperplasia through targeted vimentin to inhibit the VAV2/Rac-1 signaling pathway. [Abstract]

2025 Aug:74:609-620. PMID: 39369957

Methotrexate purchased from MedChemExpress. Usage Cited in: J Adv Res. 2025 Aug:74:609-620. [Abstract]

The cell viability of MH7A cells treated with methotrexate (1 μM) for 24 h was determined by the CCK-8 method.

Methotrexate purchased from MedChemExpress. Usage Cited in: J Adv Res. 2025 Aug:74:609-620. [Abstract]

The cell migration of MH7A cells treated with methotrexate (1 μM) for 24 h.
J Exp Clin Cancer Res

OTULIN confers cisplatin resistance in osteosarcoma by mediating GPX4 protein homeostasis to evade the mitochondrial apoptotic pathway. [Abstract]

2024 Dec 26;43(1):330. PMID: 39721999
J Nanobiotechnology

EphA2-specific microvesicles derived from tumor cells facilitate the targeted delivery of chemotherapeutic drugs for osteosarcoma therapy. [Abstract]

2024 Mar 3;22(1):89. PMID: 38433190
Small

DNA Base Pairing-Inspired Supramolecular Nanodrug Camouflaged by Cancer-Cell Membrane for Osteosarcoma Treatment. [Abstract]

2022 Jul;18(30):e2202337. PMID: 35780479
Redox Biol

Therapeutic potential of monomethyl fumarate and aluminum ion combination in alleviating inflammation and oxidative stress in psoriasis. [Abstract]

2024 Dec 25:79:103482. PMID: 39736200
Cell Rep Med

CAN-Scan: A multi-omic phenotype-driven precision oncology platform identifies prognostic biomarkers of therapy response for colorectal cancer. [Abstract]

2025 Apr 2:102053. PMID: 40187357
Mol Biomed

A single non-coding SNP in FPGS modulates folate drug efficacy in acute lymphoblastic leukemia: data-driven exploration and experimental validation. [Abstract]

2025 Nov 21;6(1):114. PMID: 41269429
Cell Death Dis

Downregulation of ZFP36L1 contributes to methotrexate resistance in osteosarcoma through enhanced NHEJ DNA repair mechanisms. [Abstract]

2025 Nov 24;16(1):852. PMID: 41285712
Cell Death Dis

Targeting TIMM23 to overcome osteosarcoma chemoresistance. [Abstract]

2025 Nov 24;16(1):856. PMID: 41285805
Acta Biomater

Impact of scaffold material choice on osteosarcoma phenotype and drug responses in 3D. [Abstract]

2025 Aug 27:S1742-7061(25)00640-3. PMID: 40882907
Biomater Res

Albumin Nanocages with Methotrexate and Chondroitin Sulfate as a Dual pH/GSH-Responsive Tumor Targeting Nanomedicine for Synergistic Cancer Therapy. [Abstract]

2025 Sep 3:29:0245. PMID: 40908968
Cell Death Dis

Deprivation of methionine inhibits osteosarcoma growth and metastasis via C1orf112-mediated regulation of mitochondrial functions. [Abstract]

2024 May 20;15(5):349. PMID: 38769167
Cell Death Dis

DEPTOR is a direct p53 target that suppresses cell growth and chemosensitivity. [Abstract]

2020 Nov 12;11(11):976. PMID: 33184290
J Pharm Anal

A highly efficient protein corona-based proteomic analysis strategy for the discovery of pharmacodynamic biomarkers. [Abstract]

2022 Dec;12(6):879-888. PMID: 36605576
Acta Pharmacol Sin

Bardoxolone displays potent activity against triple negative breast cancer by inhibiting the TRIP13/STAT3 circuit. [Abstract]

2025 Jun;46(6):1733-1741. PMID: 39939802
Acta Pharmacol Sin

Osimertinib successfully combats EGFR-negative glioblastoma cells by inhibiting the MAPK pathway. [Abstract]

2021 Jan;42(1):108-114. PMID: 32398685
Phytomedicine

Paeoniflorin derivative ameliorates methotrexate resistance in treating rheumatoid arthritis through targeting GRK2-A2AAR axis. [Abstract]

2025 Dec 19:150:157731. PMID: 41447845
EMBO Mol Med

Glycine decarboxylase advances IgA nephropathy by boosting mesangial cell proliferation through the pyrimidine pathway. [Abstract]

2025 Oct 13. PMID: 41083822
Phytomedicine

Geniposide alleviates VEGF-induced angiogenesis by inhibiting VEGFR2/PKC/ERK1/2-mediated SphK1 translocation. [Abstract]

2022 Jun;100:154068. PMID: 35358930
EMBO Mol Med

A clinically compatible drug-screening platform based on organotypic cultures identifies vulnerabilities to prevent and treat brain metastasis. [Abstract]

2022 Mar 7;14(3):e14552. PMID: 35174975
Biomed Pharmacother

Characterization of a deazaflavin analog as a potent inhibitor of multidrug resistance-associated protein 1. [Abstract]

2024 Jul 19:178:117167. PMID: 39032285
Cell Rep

Structural basis for the reversal of human MRP4-mediated multidrug resistance by lapatinib. [Abstract]

2025 Mar 25;44(4):115466. PMID: 40138312
Sci Data

High-throughput drug screening identifies novel therapeutics for Low Grade Serous Ovarian Carcinoma. [Abstract]

2024 Sep 19;11(1):1024. PMID: 39300112
Sci Signal

Coordination between the eIF2 kinase GCN2 and p53 signaling supports purine metabolism and the progression of prostate cancer. [Abstract]

2024 Nov 26;17(864):eadp1375. PMID: 39591412
Ecotoxicol Environ Saf

PPARγ-responsive luciferase reporter system for high-throughput screening of chemical toxins with potential pulmonary fibrosis effects. [Abstract]

2025 Nov 15:307:119433. PMID: 41273832
J Ethnopharmacol

Kai-Xin-San alleviates Alzheimer's disease by targeting the DHFR-mediated folate-mitochondrial axis. [Abstract]

2026 Apr 24:361:121230. PMID: 41548619
Commun Biol

Serine hydroxymethyltransferase as a potential target of antibacterial agents acting synergistically with one-carbon metabolism-related inhibitors. [Abstract]

2022 Jun 23;5(1):619. PMID: 35739195
Eur J Pharm Sci

Repurposing of FDA-approved drugs by targeting SIRT2 to alleviate inflammatory response and kidney injury. [Abstract]

2025 Sep 27:214:107296. PMID: 41022315
Int Immunopharmacol

Isorhapontigenin suppresses inflammation, proliferation and aggressiveness of rheumatoid arthritis fibroblast-like synoviocytes by targeting farnesyl diphosphate synthase. [Abstract]

2025 May 23:159:114894. PMID: 40412131
Int Immunopharmacol

Regulation of P-glycoprotein through CaMKII/cPLA2 pathway in lymphocytes for treating refractory rheumatoid arthritis by manidipine. [Abstract]

2025 Apr 26:156:114735. PMID: 40294472
Int Immunopharmacol

Cornuside alleviates psoriasis-like skin lesions in mice by relieving inflammatory effects. [Abstract]

2024 Jun 15:134:112183. PMID: 38705031
Int Immunopharmacol

Herbal compound cepharanthine attenuates inflammatory arthritis by blocking macrophage M1 polarization. [Abstract]

2023 Nov 15;125(Pt B):111175. PMID: 37976601
Eur J Pharmacol

Lycorine eliminates B-cell acute lymphoblastic leukemia cells by targeting PSAT1 through the serine/glycine metabolic pathway. [Abstract]

2023 Dec 15:961:176162. PMID: 37951487
Int Immunopharmacol

Iso-seco-tanapartholide induces p62 covalent oligomerization to activate KEAP1-NRF2 redox pathway in rheumatoid arthritis. [Abstract]

2023 Feb:115:109689. PMID: 36621330
Arthritis Res Ther

Penfluridol targets acid sphingomyelinase to inhibit TNF signaling and is therapeutic against inflammatory autoimmune diseases. [Abstract]

2022 Jan 19;24(1):27. PMID: 35045889
Toxicology

Drug interaction study of flavonoids toward OATP1B1 and their 3D structure activity relationship analysis for predicting hepatoprotective effects. [Abstract]

2020 May 15;437:152445. PMID: 32259555
Cell Rep Methods

RECOVER identifies synergistic drug combinations in vitro through sequential model optimization. [Abstract]

2023 Oct 23;3(10):100599. PMID: 37797618
Rheumatology (Oxford)

Autophagy inhibitors block pathogenic NET release in immune-mediated inflammatory disease without impairing host defence. [Abstract]

2025 Aug 13:keaf437. PMID: 40802538
Cancers (Basel)

Association between Dysfunction of the Nucleolar Stress Response and Multidrug Resistance in Pediatric Acute Lymphoblastic Leukemia. [Abstract]

2022 Oct 19;14(20):5127. PMID: 36291909
Cancers

Synergism Through WEE1 and CHK1 Inhibition in Acute Lymphoblastic Leukemia. [Abstract]

2019 Oct 25;11(11):1654. PMID: 31717700
ACS Omega

ROS-Scavenging Bamboo-Derived Carbon Dot-Methotrexate Nanocomposite Ameliorates Rheumatoid Arthritis through Dual Therapeutic Mechanisms. [Abstract]

2026 Jan 21;11(4):5853-5864. PMID: 41658163
ACS Omega

Association Study of OATP1B3 Polymorphisms on Hepatic Uptake and Drug-Drug Interaction In Vitro. [Abstract]

2025 Oct 29;10(44):52562-52575. PMID: 41244417
J Cell Mol Med

Inactivation of CDK12 Enhances Mitochondrial Efficiency to Suppress DNA Damage. [Abstract]

2026 Apr;30(7):e71101. PMID: 41896195
Mediators Inflamm

Peptide BG From Bitter Gourd (Momordica Charantia) Improves Adjuvant-Induced Arthritis by Modulating the Necroptosis/Neutrophil Extracellular Traps/Inflammation Axis and the Gut Microbiota. [Abstract]

2024 Dec 5:2024:1995952. PMID: 39669913
J Mol Med (Berl)

Combination of PARP inhibitor and temozolomide to suppress chordoma progression. [Abstract]

2019 Aug;97(8):1183-1193. PMID: 31201471
Lung

Disulfiram Alleviates MTX-Induced Pulmonary Fibrosis by Inhibiting EMT in Type 2 Alveolar Epithelial Cells. [Abstract]

2024 Nov 27;203(1):4. PMID: 39601871
Sci Rep

High expression of LncRNA HOTAIR is a risk factor for temozolomide resistance in glioblastoma via activation of the miR-214/β-catenin/MGMT pathway. [Abstract]

2024 Oct 31;14(1):26224. PMID: 39482401
J Biol Chem

A technique for delineating the unfolding requirements for substrate entry into retrotranslocons during endoplasmic reticulum-associated degradation. [Abstract]

2019 Dec 27;294(52):20084-20096. PMID: 31748412
Sci Rep

A widely-applicable high-throughput cellular thermal shift assay (CETSA) using split Nano Luciferase. [Abstract]

2018 Jun 21;8(1):9472. PMID: 29930256
Chem Res Toxicol

Inhibitory Effects of Alkaloids on OATP1B1 In Vitro and In Vivo: Prediction for Food/Herb-Drug Interactions and Hepatoprotective Effects Based on Structure-Activity Relationships. [Abstract]

2025 Feb 17;38(2):281-295. PMID: 39899883
Biotechnol Bioeng

An integrated biomimetic array chip for establishment of collagen-based 3D primary human hepatocyte model for prediction of clinical drug-induced liver injury. [Abstract]

2021 Dec;118(12):4687-4698. PMID: 34478150
J Bone Oncol

Identification of GPC3 mutation and upregulation in a multidrug resistant osteosarcoma and its spheroids as therapeutic target. [Abstract]

2021 Sep 20:30:100391. PMID: 34611509
Dis Model Mech

A Drosophila chemical screen reveals targeting MEK and DGKa mitigates Ras-driven polarity-impaired tumour growth. [Abstract]

2023 Mar 1;16(3):dmm049769. PMID: 36861754
Mol Immunol

Methotrexate loaded extracellular vesicles attenuate periodontitis by suppressing ACSL1 and promoting anti-inflammatory macrophage. [Abstract]

2025 Apr 16:182:83-95. PMID: 40245705
Micromachines

A Novel SimpleDrop Chip for 3D Spheroid Formation and Anti-Cancer Drug Assay. [Abstract]

2021 Jun 10;12(6):681. PMID: 34200752
Plasma Process Polym

2021 Feb 12.
Int J Immunopathol Pharmacol

Cucurbitacin B inhibits Th17 cell differentiation via the suppression of the JAK/STAT pathway and alleviates collagen-induced arthritis in mice. [Abstract]

2025 Jan-Dec:39:3946320251348715. PMID: 40518910
Biochem Biophys Rep

Histone H3 lysine-trimethylation markers are decreased by recombinant methioninase and increased by methotrexate at concentrations which inhibit methionine-addicted osteosarcoma cell proliferation. [Abstract]

2021 Nov 26:28:101177. PMID: 34877414
Anticancer Res

Loss of Malignancy of Super-Methotrexate-resistant Osteosarcoma Cells Is Associated With an Increase of Methylated Histone Marks H3K9me3 and H3K27me3. [Abstract]

2024 Oct;44(10):4213-4218. PMID: 39348992
Anticancer Res

Reduced Malignancy of Super Methotrexate-resistant Osteosarcoma Cells With Dihydrofolate Reductase Amplification Despite Paradoxical Gain of Oncogenic PI3K/AKT/mTOR and c-MYC expression. [Abstract]

2024 Jul;44(7):2787-2792. PMID: 38925854
bioRxiv

2026 May 6.
bioRxiv

2025 Nov 20.
SSRN

2025 Oct 10.
Patent

US20250235423A1.
Authorea

2025 Jan 03.
Guidelines and Standards in Chinese Medicine

2024 Dec.
Research Square Preprint

2023 Dec 4.
Research Square Preprint

2023 Dec 1.
Queen’s University

2021 Oct.
Oncotarget

Molecular-genetic profiling and high-throughput in vitro drug screening in NUT midline carcinoma-an aggressive and fatal disease. [Abstract]

2017 Dec 2;8(68):112313-112329. PMID: 29348827
Patent

US20170128439A1.

Solvent & Solubility

In Vitro:

DMSO : ≥ 50 mg/mL (110.03 mM; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

* "≥" means soluble, but saturation unknown.

View the Complete Stock Solution Preparation Table

Preparing Stock Solutions

View the Complete Stock Solution Preparation Table

Concentration Solvent Mass

1 mg

5 mg

10 mg

Concentration

Solvent

Mass

1 mM

2.2005 mL

11.0026 mL

22.0051 mL

5 mM

0.4401 mL

2.2005 mL

4.4010 mL

10 mM

0.2201 mL

1.1003 mL

2.2005 mL

Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.

Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

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In Vivo:

Select the appropriate dissolution method based on your experimental animal and administration route.

Stock Solution Working Solution

For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for In Vivo experiments, it is recommended to prepare freshly and use it on the same day.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

Protocol 1

Add each solvent one by one: 10% DMSO 40% PEG300 5% Tween-80 45% Saline

Solubility: ≥ 2.5 mg/mL (5.50 mM); Clear solution

This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.

Protocol 2

Add each solvent one by one: 10% DMSO 90% (20% SBE-β-CD in Saline)

Solubility: ≥ 2.5 mg/mL (5.50 mM); Clear solution

This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.

Protocol 3

Add each solvent one by one: 5% DMSO 40% PEG300 5% Tween-80 50% Saline

Solubility: ≥ 2.5 mg/mL (5.50 mM); Clear solution

Protocol 4

Add each solvent one by one: 5% DMSO 95% (20% SBE-β-CD in Saline)

Solubility: ≥ 2.5 mg/mL (5.50 mM); Clear solution

In Vivo Dissolution Calculator

Please enter the basic information of animal experiments:

Dosage Dosage
mg/kg
Animal Weight (Per Animal) Animal Weight (Per Animal)
g
Dosing Volume (Per Animal) Dosing Volume (Per Animal)
μL
Number of Animals Number of Animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.

Please enter your animal formula composition:

%
DMSO +

Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
%
+
+
%
Tween-80 + +
%
Saline +

The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).

Working solution concentration: 0.22 mg/mL

Protocol

Cell Assay^[1]

Each cell line is studied in growth inhibition experiments using 96-well microtiter plates. As antifols are schedule dependent, preliminary experiments are aimed at defining the longest duration of exposure that would allow for continuous logarithmic phase growth of cells without changing of the culture media while maintaining a linear relationship between SRB optical density and cell number. Twenty-four hours after cell plating, the cell lines are exposed to the antifol for 120 h (three replicates per experiment). To ensure that a complete sigmoidal survival-concentration curve could be observed, the following drug concentrations are studied: Methotrexate (0.002-5 μM), AMT (0.0001-1 μM), PXD (0.0003-10 μM), TLX (0.0002-0.5 μM). Experiments are repeated at least twice^[1].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Animal Administration^[2]

Mice^[2]
The combination of bioactive phytochemicals is administered one week prior to the Methotrexate exposure. Treatment group I: mice are given a combination of green tea polyphenols and eleutherosides from Siberian ginseng (0.2 mL/10 g, i.g. once daily) for 15 days, and a single dose of Methotrexate (2 mg/kg, i.p. once daily) is added on the 8th day. Treatment group II: mice are given a combination of grape seed proanthocyanidins and eleutherosides from Siberian ginseng for 15 days, and Methotrexate is administered on the 8th day in a similar manner. Model group: animals received distilled water instead of bioactive phytochemicals combinations for 15 days and the same Methotrexate protocol applied to this group on the 8th day. Control group: mice are given distilled water through 15 days and physiological saline instead of Methotrexate is administered on the 8th day in a similar manner. Twelve hours after the final doses, the animals are euthanized by cervical dislocation.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Purity & Documentation

Select Batch:

Purity: 99.98%

Assay: 99.98%

ee.: 99.98%

Data Sheet (277 KB)
SDS (644 KB)
- English - EN (644 KB)
- Français - FR (644 KB)
- Deutsch - DE (644 KB)
- Norwegian - NO (644 KB)
- Español - ES (644 KB)
- Swedish - SV (644 KB)
- Italian - IT (644 KB)
- Korean - KR (644 KB)
- Portuguese - PT (644 KB)
COA (2659 KB)
Elemental Analysis Report (2659 KB)
HNMR (2659 KB)
CNMR (2659 KB)
RP-HPLC (2659 KB)
NP-HPLC (2659 KB)
MS (2659 KB)
LCMS (2659 KB)
IR (2659 KB)
GC (2659 KB)
OR (2659 KB)
SDS-PAGE (2659 KB)
SEC-HPLC (2659 KB)
Other Report (2659 KB)
Handling Instructions (2659 KB)

References

[1]. Tian H, et al. Understanding the mechanisms of action of methotrexate: implications for the treatment of rheumatoid arthritis. Bull NYU Hosp Jt Dis. 2007;65(3):168-73. [Content Brief]

[2]. Swierkot J, et al. Methotrexate in rheumatoid arthritis. Pharmacol Rep. 2006 Jul-Aug;58(4):473-92. [Content Brief]

[3]. Ehab Tousson, et al. The Effect of L-carnitine on Amethopterin-induced Toxicity in Rat Large Intestine.

[4]. Banji D, et al. Evaluation of the concomitant use of methotrexate and curcumin on Freund's complete adjuvant-induced arthritis and hematological indices in rats. Indian J Pharmacol. 2011;43(5):546-550. [Content Brief]

Help & FAQs

How should lyophilized recombinant proteins be reconstituted and stored?

1. Before opening the cap, centrifuge the vial at 13000 rpm for 20-30 seconds. This step will ensure that any lyophilized powder that may have adhered to the cap or walls is collected at the bottom of the vial, minimizing the risk of product loss.

Complete Stock Solution Preparation Table

Optional Solvent	Concentration Solvent Mass	1 mg	5 mg	10 mg	25 mg
DMSO	1 mM	2.2005 mL	11.0026 mL	22.0051 mL	55.0128 mL
	5 mM	0.4401 mL	2.2005 mL	4.4010 mL	11.0026 mL
	10 mM	0.2201 mL	1.1003 mL	2.2005 mL	5.5013 mL
	15 mM	0.1467 mL	0.7335 mL	1.4670 mL	3.6675 mL
	20 mM	0.1100 mL	0.5501 mL	1.1003 mL	2.7506 mL
	25 mM	0.0880 mL	0.4401 mL	0.8802 mL	2.2005 mL
	30 mM	0.0734 mL	0.3668 mL	0.7335 mL	1.8338 mL
	40 mM	0.0550 mL	0.2751 mL	0.5501 mL	1.3753 mL
	50 mM	0.0440 mL	0.2201 mL	0.4401 mL	1.1003 mL
	60 mM	0.0367 mL	0.1834 mL	0.3668 mL	0.9169 mL
	80 mM	0.0275 mL	0.1375 mL	0.2751 mL	0.6877 mL
	100 mM	0.0220 mL	0.1100 mL	0.2201 mL	0.5501 mL

Methotrexate

77 Cited Publications

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Methotrexate

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Chemical Information

Publications (77)

Solvent & Solubility

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Complete Stock Solution Preparation Table

Methotrexate Related Classifications

Cat. No.: HY-14519