Anlotinib Overcomes Multiple Drug Resistant Colorectal Cancer Cells via Inactivating PI3K/AKT Pathway

Background: Anlotinib is a multi-target tyrosine kinase inhibitor that has been reported to have activity against colorectal Cancer. However, the mechanisms of how anlotinib mediates drug-resistance of colorectal Cancer have not been fully described. Particularly the potential mechanisms regarding the inhibition of proliferation and induction of Apoptosis remain unknown.

Objective: In this study, we intended to study the effect and related-mechanism of the proliferation, migration, invasion and induced Apoptosis of anlotinib overcoming multidrug resistant colorectal Cancer cells through in vitro experiments.

Methods: Cell viability was determined by MTT assays and the resistant index was calculated. Colony formation and PI/RNase Staining were used for testing the proliferation of resistant cells. DAPI staining and Annexin V-FITC/PI staining were used to detect cell Apoptosis. Migration and invasion were examined by transwell. Protein expression and activation of PI3K/Akt pathway were detected by western blot. LY294002 was used to verify whether anlotinib overcomes the drug-resistance of CRC cells by inactivating the PI3K/Akt pathway.

Results: The results showed that the HCT-8/5-FU cells were resistant to multiple chemotherapy drugs (5-FU, ADM and DDP). Anlotinib significantly inhibited cell viability, proliferation, migration, invasion and induced cell Apoptosis. Moreover, anlotinib down-regulated the expression of Survivin, cyclin D1, CDK4, Caspase-3, Bcl-2, MMP-2, MMP-9, vimentin and N-Cadherin, but up-regulated cleaved-caspase-3, Bax and E-cadherin and blocked the activity of the PI3K/Akt in HCT-8/5-FU cells. We found anlotinib and LY294002 overcame the drug resistance of HCT-8/5-FU cells by reducing the expression of PI3K/p-AKT.

Conclusion: Anlotinib inhibited the proliferation, migration, invasion and induced Apoptosis of HCT-8/5-FU cells, and the mechanisms may be that anlotinib conquered multidrug resistance of colorectal Cancer cells via inactivating of PI3K/Akt pathway.