2. Apoptosis MAPK/ERK Pathway Metabolic Enzyme/Protease Immunology/Inflammation NF-κB
  3. MDM-2/p53 RIP kinase Mixed Lineage Kinase Reactive Oxygen Species (ROS) Necroptosis
  4. RETRA

RETRA is a RIPK1/RIPK3/MLKL and p73 activator. RETRA mediates plasma membrane disruption, induces ROS accumulation, triggers early mitochondrial hyperpolarization, and synergistically drives necroptosis. RETRA inhibits cancer cells carrying mutant p53 via a p73-dependent salvage pathway. RETRA can be used in research related to cervical cancer and cancers with mutant p53.

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RETRA

RETRA Chemical Structure

CAS No. : 1036069-26-7

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Description

RETRA is a RIPK1/RIPK3/MLKL and p73 activator. RETRA mediates plasma membrane disruption, induces ROS accumulation, triggers early mitochondrial hyperpolarization, and synergistically drives necroptosis. RETRA inhibits cancer cells carrying mutant p53 via a p73-dependent salvage pathway. RETRA can be used in research related to cervical cancer and cancers with mutant p53[1][2].

In Vitro

RETRA (10-100 μM; 24-96 h) inhibits the viability of SiHa and C-33A cervical cancer cells in a concentration- and time-dependent manner[1].
RETRA (25-100 μM; 24-72 h) inhibits the proliferation of SiHa and C-33A cervical cancer cells in a concentration-dependent manner[1].
RETRA (25-100 μM; 48 h) induces S-phase cell cycle arrest in SiHa and C-33A cervical cancer cells by regulating cell cycle regulatory proteins, including upregulating p21 and downregulating cyclin D3[1].
RETRA (25-100 μM; 24, 48, 72 h) selectively induces non-apoptotic cell death in SiHa and C-33A cervical cancer cells, and exhibits no cytotoxicity against normal human peripheral blood mononuclear cells (PBMCs)[1].
RETRA (50-100 μM; 48 h) induces necroptosis in SiHa, C-33A and CaSki cervical cancer cells via phosphorylation of RIPK1, RIPK3 and MLKL; this cell death is reversible by the necroptosis inhibitor Nec-1 (HY-15760)[1].
RETRA (25-100 μM; 24, 48 h) induces mitochondrial hyperpolarization in SiHa and C-33A cervical cancer cells in a concentration- and time-dependent manner[1].
RETRA (25-100 μM; 48 h) induces concentration-dependent ROS accumulation in SiHa and C-33A cervical cancer cells, and this accumulation is attenuated by the necroptosis inhibitor Nec-1[1].
RETRA (1-10 μM; 14 h) specifically activates p53-dependent transcription reporter gene activity in human cancer cell lines expressing mutant p53 (including A431, SW480 and MDA-MB-231) as well as p53-deficient cell lines reconstituted with mutant p53, but exerts no effect on cells expressing wild-type p53 or p53-deficient cells[2].
RETRA (1.5 μg/mL; 14 h) selectively induces transcriptional activation of the p53-dependent genes CDKN1A and BBC3 in A431 cells expressing mutant p53[2].
RETRA (2 μg/mL; 20 h) increases TA/p73 protein levels in A431 cells expressing mutant p53 and releases TA/p73 from the inhibitory complex formed with mutant p53; whereas after treatment at 2 μg/mL for 20 h[2].
RETRA (0.1-10 μM; 48 h) induces dose-dependent inhibition of cell viability in A431 cells expressing mutant p53, and exhibits enhanced activity in p53-knockdown A431/sh-p53 cells[2].
RETRA (4 μM; 24 h) specifically inhibits colony formation of A431 and SW480 cells expressing mutant p53, and this effect is partially dependent on the expression of p73; while after treatment with 4 μM for 24 h[2].
RETRA (1-10 μM) induces dose-dependent activation of caspase 3 and caspase 7 in A431 cells expressing mutant p53, and the activity is enhanced in p53-knockdown A431/sh-p53 cells[2].

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

RETRA Related Antibodies

Cell Viability Assay[1]

Cell Line: SiHa (wild-type p53 cervical cancer), C-33A (mutant p53 cervical cancer) cells
Concentration: 10, 20, 30, 40, 50, 60, 70, 80, 90 and 100 μM
Incubation Time: 24, 48, 72, 96 h
Result: Elicited a concentration- and time-dependent decrease in cell viability in both SiHa and C-33A cells, with significant reductions observed at higher concentrations and longer incubation times.

Cell Proliferation Assay[1]

Cell Line: SiHa (wild-type p53 cervical cancer), C-33A (mutant p53 cervical cancer) cells
Concentration: 25, 50, 75 and 100 μM
Incubation Time: 24, 48, 72 h
Result: Resulted in significantly higher residual CFSE fluorescence compared to untreated cells, indicating reduced cell proliferation in a concentration-dependent manner.
Showed a dramatic reduction in cell numbers and altered morphology at 72 h of treatment via phase-contrast microscopy.

Cell Cycle Analysis[1]

Cell Line: SiHa (wild-type p53 cervical cancer), C-33A (mutant p53 cervical cancer) cells
Concentration: 25, 50, 75 and 100 μM
Incubation Time: 48 h; 96 h (sub-G1 analysis)
Result: Caused a substantial accumulation of cells in the S-phase (approximately 47%) after 48 h of treatment, along with a concentration-dependent increase in sub-G1 phase cells (up to 33% in C-33A at 96 h).
Decreased cyclin D3 levels, increased p21 and CDK4 levels, and increased p73 levels in SiHa cells, with no change in p53 levels via Western blot analysis.

Apoptosis Analysis[1]

Cell Line: SiHa (wild-type p53 cervical cancer), C-33A (mutant p53 cervical cancer) cells, normal human peripheral blood mononuclear cells (PBMCs)
Concentration: 25, 50, 75 and 100 μM
Incubation Time: 24, 48, 72 h (cancer cells); 24, 48, 72 h (PBMCs)
Result: Caused a concentration- and time-dependent increase in Annexin V-/PI+ (necrotic/necroptotic) cells in SiHa and C-33A cells, with no significant increase in apoptotic cell populations and no change in cleaved PARP-1 levels.
Showed no significant increase in apoptotic or necrotic cell populations in PBMCs even at the highest concentration and longest incubation time.

Western Blot Analysis[2]

Cell Line: A431, H1299, A549
Concentration: 2 μg/mL
Incubation Time: 20 h
Result: Induced a 4- to 5-fold increase in TA/p73 protein levels in A431 cells.
Caused no increase in TA/p73 levels in p53-null H1299 or wild-type p53-expressing A549 cells.
Reduced the fraction of TA/p73 co-immunoprecipitated with mutant p53 by 2- to 3-fold in A431 cells, leading to an ~10-fold increase in free TA/p73 compared to untreated controls.

Cell Viability Assay[2]

Cell Line: A431, A431/sh-p53, A431/sh-p73
Concentration: 0.1, 1 and 10 μM
Incubation Time: 48 h
Result: Caused dose-dependent inhibition of cell viability in A431 cells.
Showed enhanced inhibition of cell viability in A431/sh-p53 cells.
Had significantly reduced viability-inhibiting effect in A431/sh-p73 cells at all tested concentrations.

Cell Proliferation Assay[2]

Cell Line: A431, A431/sh-p73, SW480, A549, H1299, PC3
Concentration: 4 μM
Incubation Time: 24 h
Result: Dramatically reduced colony formation in A431 and SW480 cells.
Had a much milder effect on colony formation in A431/sh-p73 cells.
Showed almost no effect on colony formation in A549, H1299, or PC3 cells.
In Vivo

RETRA (0.4 mg/mouse; i.p.; daily; 6 days) suppresses A431 xenograft tumor formation in athymic nu/nu mice, reducing the percentage of tumor-positive injection sites to ~40%[2].

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

Animal Model: athymic nu/nu mice[2]
Dosage: 0.4 mg/mouse
Administration: i.p.; daily; 6 days
Result: Delayed tumor formation time.
Reduced the final percentage of tumor-forming injection sites to ~40%.
Molecular Weight

269.34

Formula

C11H11NO3S2
CAS No.
Appearance

Solid

Color

Off-white to gray

SMILES

OC1=CC=C(C(CSC2=NCCS2)=O)C=C1O
Shipping

Room temperature in continental US; may vary elsewhere.
Storage

-20°C, protect from light

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

Solvent & Solubility
In Vitro: 

DMSO : 125 mg/mL (464.10 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

Preparing
Stock Solutions
Concentration Solvent Mass 1 mg 5 mg 10 mg
1 mM 3.7128 mL 18.5639 mL 37.1278 mL
5 mM 0.7426 mL 3.7128 mL 7.4256 mL
View the Complete Stock Solution Preparation Table

* 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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Purity & Documentation
References

Complete Stock Solution Preparation Table

* 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.

Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
DMSO 1 mM 3.7128 mL 18.5639 mL 37.1278 mL 92.8195 mL
5 mM 0.7426 mL 3.7128 mL 7.4256 mL 18.5639 mL
10 mM 0.3713 mL 1.8564 mL 3.7128 mL 9.2819 mL
15 mM 0.2475 mL 1.2376 mL 2.4752 mL 6.1880 mL
20 mM 0.1856 mL 0.9282 mL 1.8564 mL 4.6410 mL
25 mM 0.1485 mL 0.7426 mL 1.4851 mL 3.7128 mL
30 mM 0.1238 mL 0.6188 mL 1.2376 mL 3.0940 mL
40 mM 0.0928 mL 0.4641 mL 0.9282 mL 2.3205 mL
50 mM 0.0743 mL 0.3713 mL 0.7426 mL 1.8564 mL
60 mM 0.0619 mL 0.3094 mL 0.6188 mL 1.5470 mL
80 mM 0.0464 mL 0.2320 mL 0.4641 mL 1.1602 mL
100 mM 0.0371 mL 0.1856 mL 0.3713 mL 0.9282 mL
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  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

Keywords:

RETRA1036069-26-7MDM-2/p53RIP kinaseMixed Lineage KinaseReactive Oxygen Species (ROS)NecroptosisReceptor-interacting protein kinasesRIPKMLKsA431necroptosisSW480cervical cancer cellsPBMCsMDA-MB-231SiHamutant p53C-33ACaSkiInhibitorinhibitorinhibit

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