厄罗替尼对肺腺癌细胞凋亡的影响

Effects of Erlotinib on apoptosis in human pulmonary adenocarcinoma

目的研究厄罗替尼对人肺腺癌细胞株A549细胞凋亡的影响以及对表皮生长因子受体(EGFR)蛋白表达的影响。方法采用MTT法检测不同浓度的厄罗替尼对A549细胞的增殖抑制作用;以倒置相差显微镜、流式细胞术、TUNEL法及DAPI荧光染色法检测细胞凋亡;以免疫荧光法分析厄罗替尼对EGFR蛋白表达的影响。结果厄罗替尼抑制A549生长呈时间浓度依赖性;厄罗替尼处理A549后倒置相差显微镜、DAPI及TUNEL均见到明显的凋亡细胞(P<0.05);药物处理组使细胞发生明显的G1期阻滞(P<0.05);TUNEL检测到200μmol/L厄罗替尼组细胞凋亡率为(42.13±1.4)%,显著高于对照组(0.82±0.03)%(P<0.05);免疫荧光法表明厄罗替尼明显下调EGFR表达(P<0.05)。结论厄罗替尼杀伤A549细胞的机制与介导细胞凋亡、阻滞细胞周期于G/G期及阻滞EGFR信号途径有关。

Objective To explore the effects of Erlotinib on apoptosis andexpression of EGFR in human pulmonary adenocarcinoma cell line A549 cells.Methods MTT assay was used to determine A549 cells proliferation.Inverted microscope,flow cytometry,DAPI and TUNEL were used to detect apoptosis.The expression of EGFR was determined by immunofluorescence.Results Erlotinib inhibited the growth of A549 cells in a dose-dependent and time-dependent.The cells treated with Erlotinib showed a typical apoptotic morphology （P0.05） and G1 phase arrestting（P0.05）.The apoptosis rate of 200 μmol/L Erlotinib group 〔（42.13±1.4）%〕 was significantly higher than that of control group〔（0.82±0.03）%〕 （P0.05）.Immunofluorescence assay showed that Erlotinib down-regulated the expression of EGFR.Conclusions Erlotinib can kill A549 cells,owing to inducing apoptosis and G1 phase arresting and down-regulating the expresion of EGFR.