盐酸埃克替尼诱导肺癌HCC827细胞周期阻滞及其机制研究

目的 研究国家一类新药盐酸埃克替尼（Icotinib）对人非小细胞肺癌细胞HCC827的增殖抑制及细胞周期的影响,并探讨其作用机制.方法 实验分为空白组、对照组和Icotinib处理组,采用四甲基偶氮唑盐法（MTT）检测Icotinib对人肺癌HCC827细胞增殖的影响；流式细胞仪检测细胞周期变化；Western blot检测相关蛋白表达,应用SPSS 13.0进行统计学分析.结果 Icotinib以时间-剂量依赖的方式抑制HCC827细胞增殖,48 h的IC50为0.60 μmol/L,72 h的IC50为0.06 μmol/L;Icotinib诱导HCC827细胞发生明显的G1期阻滞并具有剂量依赖性.进一步对周期相关蛋白检测发现,Icotinib处理组较对照组相比,显著上调p21蛋白表达,抑制cyclin D1及cyclin A表达,但对cyclin E作用不明显.检测还发现Icotinib处理组明显下调ERK的磷酸化水平.结论 Icotinib能够明显抑制HCC827细胞增殖,引起细胞G1期阻滞,其机制可能与上调p21及抑制cyclin D1、cyclin A蛋白表达水平相关.并且MAPK/ERK信号通路在其介导的生物学效应中起重要作用.

Effects of icotinib on cell cycle arrest in human lung cancer HCC827 cells

Objective To explore the effects of icotinib on the cell proliferation and cell cycle in human non-small cell lung cancer （NSCLC） HCC827 cells,and to further investigate the mechanism of action.Methods Cell proliferation was measured by using MTT assay.Cell cycle changes were determined by flow cytometry.The expressions of proteins were detected by Western blot.All experimental data were dealt with SPSS （13.0 soft）.Results Icotinib significantly inhibited HCC827 cell viability in a concentration and time dependent manner,the concentration of inhibited cell viability （IC50） for 48 h was 0.60 μmol/L,72 h was 0.06 μmol/L.Icotinib induced significant G1 phase arrest of HCC827 cells in a concentration-dependent manner.Further more,icotinib could upregulated the expression of p21,decreased the expression of cyclin D1 and cyclin A,but had no effect on cyclin E.In addition,after treated 24 h,icotinib could inhibit the expressions of the phosphorylated ERK obviously in HCC827 cells.Conclusions Icotinib inhibited the activation of the ERK signaling pathway,which consequently upregulated the expression of p21,downregulated cyclin D1 and cyclin A in HCC827 cells.Furthermore,MAPK/ERK pathway plays an important role in the biological effects of icotinib.