塞来昔布对埃克替尼诱导肺腺癌细胞A549凋亡作用的影响

目的 研究埃克替尼与塞来昔布联合应用对人肺腺癌A549细胞凋亡的影响以及对EGFR、COX-2表达的影响。方法 埃克替尼、塞来昔布单独或联合干预细胞48h后,倒置相差显微镜观察药物干预后细胞形态学变化;四甲基偶氮唑盐（MTT）比色法测定细胞抑制率;HOECHEST33258荧光染色法和流式细胞术检测细胞凋亡和药物作用周期;免疫荧光检测EGFR和COX-2蛋白表达情况。结果 埃克替尼联合塞来昔布,相比单药组明显出现大量颗粒和空泡,细胞变圆开始脱落;埃克替尼与塞来昔布对A549细胞增殖有明显抑制作用,并呈浓度及时间依赖性,二者联合作用时抑制作用更强（P〈0.05）;埃克替尼与塞来昔布均能诱导细胞凋亡,联合作用后细胞凋亡率更高（P〈0.05）;联合用药与单独用药相比,可使细胞发生明显的G1期阻滞（P〈0.05）,进一步下调了EGFR和COX-2蛋白的表达（P〈0.05）。结论 埃克替尼与塞来昔布联合应用具有协同作用,机制可能与介导细胞凋亡,阻滞细胞周期于G0/G1期及阻滞EGFR和COX-2信号途径有关。

EFFECT OF CELECOXIB ON THE APOPTOSIS INDUCED BY ICOTINIB IN LUNG CANCER A 549 CELLS

Objective To investigate the effects of Icotinib combined with Celecoxib on apoptosis and expression of EGFR and COX-2in lung cancer A549 cells.Methods A549 cells were treated with Icotinib,Celecoxib alone or in combination for 48 hours.Cellular morphological changes were observed under an inverted microscope,MTT was used to measure the growth inhibitory effects of A549 cells by Icotinib and Celecoxib.The cell apoptosis and cell cycle were detected by HOECHEST33258 staining method and flow hyetometer.The expression of EGFR,COX-2 was determined by immunofluorescence.Results A great quantity of granules and vacuoles were observed in the combined treatment group,with cell rounding and defluxion.Icotinib and Celecoxib inhibited the growth of A549 cells in a dose- dependent and time-dependent.Their combination produced greater growth inhibition than used alone（P〈0.05）.Compared with either Icotinib or Celecoxib alone,combination increased apoptosis of A549（P〈0.05）significantly.Icotinib and Celecoxib also induced G1 phase arrest and down-regulated expression of EGFR,COX-2 in A549cells（P〈0.05）.Conclusion The results suggest that combination treatment with Icotinib and Celecoxib can improve therapeutic efficacy against A549 cells,owing to inducing apoptosis,G1 phase arrest,and down-regulated expressing of both EGFR and COX-2.