【摘要】 目的:探讨酪氨酸激酶抑制剂（tyrosine kinase inhibitor,TKI）阿法替尼（afatinib）对乳腺癌细胞增殖、周期及凋亡的影响,并就阿法替尼与吉非替尼（gefitinib）对乳腺癌细胞的作用进行比较。方法:应用MTT法检测人乳腺癌细胞系MCF-7、T47D、MDAMB-231细胞活性,流式细胞术的PI染色法检测细胞周期变化以及Annexin-V/PI双染法检测细胞凋亡,通过Western blot法检测蛋白表达情况。结果:阿法替尼对MCF-7、T47D、MDA-MB-231细胞均有明显的抑制作用,IC₅₀分别为0.101、0.141、0.887μmol/L。阿法替尼对T47D、MDA-MB-231细胞作用24 h后G₀/G₁期细胞比例明显升高,细胞凋亡率增加,晚期的凋亡率分别为88.9%、58.1%,并可促进细胞凋亡通路蛋白PARP、caspase-3发生剪切。阿法替尼、吉非替尼使MDA-MB-231细胞的EGFR磷酸化水平受到明显抑制,相同浓度下,阿法替尼作用较吉非替尼更强、持续时间更长。结论:阿法替尼可显著抑制乳腺癌细胞增殖、促进其凋亡,且具有明显的量效关系,较吉非替尼具有更有效的作用。更多还原

【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.更多还原