吉非替尼对A549细胞增殖及细胞周期、凋亡的作用

目的：观察吉非替尼对人非小细胞肺癌A549的生长增殖、细胞周期及细胞凋亡的影响。方法：用5—40μmol／L浓度范围内的吉非替尼和A549细胞共培养24、48、72h，采用四甲基偶氮唑蓝（MTr）法检测吉非替尼对细胞增殖的影响；用流式细胞仪检测其对细胞凋亡及细胞周期分布的影响；利用抗Capspase-3的ELISA（酶联免疫吸附）法检测是否发生细胞凋亡。结果：在5—40μmol／L浓度范围内，吉非替尼对A549细胞的增殖有明显的抑制作用，并呈时间和剂量依赖性，以40μmol／L作用72h时的抑制率最高。流式细胞仪检测显示10μmol／L-401μmol／L的吉非替尼作用可使细胞发生G0／G，期阻滞，但即使作用48h也未发现凋亡细胞。ELISA法显示48h内，吉非替尼组与正常细胞组的Capspase-3浓度无统计学差异，但作用72h时出现凋亡。结论：吉非替尼在5—40μmol／L浓度下作用时间小于48h时，其对A549细胞的增殖抑制可能是通过阻滞A549细胞于G0／G1期而非引起细胞凋亡实现的，但作用时间达到72h时，其可以诱导A549发生细胞凋亡。

Effect of gefitinib on proliferation and cell cycle, apoptosis of A549 cells

Objective：To investigate the effect of gefitinib on the proliferation and cell cycle, apoptosis in non - small cell lung cancer A549 cells. Methods：AS49 cells were exposed to different concentration of gefitinib for 24h, 48h and 72h respectively. The proliferation was detected by 4 -methyl -tetrazolium （MTT） ,the distribution of cell cycle and effect on apoptosis of A549 cells exposed to different concentration of gefitinib was analyzed by flow cytome- try and anti - Capspase - 3 ELISA kit. Results： Within the range of 5 - dO p~mol/L, gefitinib significantly inhibited the proliferation of A549 cells in a dose - and time - dependent manner. 10 ~xmol/L - 40 p~mol/L gefitinib caused A549 cells to undergo G0/G1 arrest but didn＇t induce apoptosis even though the action time to achieve 48h. The result of ELISA showed A549 cells cocultured with gefitinib for 24h,48h didn＇t happen apeptosis,but not the cells for 72h. Conelusion：Gefitinib which do effect within the range of 5 -40 ~mol/L for 24,48h could inhibit the proliferation of A549 cells by blocking the cell cycle in G0/G1 phase while not induce apoptosis. When gefitinib cocultured with A549 cells for 72h,it induced apoptosis.