Abstract:
Objective To investigate the effect of afatinib, a tyrosine kinase inhibitor, on the proliferation, cell cycle, and apoptosis of human breast cell lines, and compare its effects with those of gefitinib.
Methods Three human breast cell lines, MCF-7, T47D, and MDA-MB-231, were cultured as cell models. A methyl thiazolyl tetrazolium assay was utilized to measure cell viability. Flow cytometer was used to analyze the cell cycle arrest (PI staining) and apoptosis rates (Annexin-V/PI staining). The protein expression was detected by Western blot analysis.
Results The proliferation of three human breast cell lines was significantly inhibited by afatinib, and the IC50 levels of MCF-7, T47D, and MDA-MB-231 were 0.101, 0.141, and 0.887 μmol/L, respectively. The G0/G1 phase cell ratio increased considerably 24 h after afatinib was added to T47D or MDA-MB-231. The cell apoptosis rate also increased in the two cell lines (88.9% and 58.1%). The cleavage of apoptosis pathway proteins PARP and caspase-3 was also promoted by afatinib. Phosphorylation of EGFR was significantly inhibited by afatinib in the MDA-MB-231 cell line. Finally, the inhibition effect of afatinib was stronger than that of gefitinib.
Conclusion Afatinib could significantly inhibit the proliferation of breast cancer cells and promote apoptosis. The effect was dose-dependent. Afatinib was a more effective tyrosine kinase inhibitor as compared with gefitinib.
下载: