厄罗替尼联合塞来昔布阻断EGFR和COX-2抑制肺癌A549细胞增殖

Erlotinib combined with celecoxib inhibits the growth of lung cancer A549 cells by simultaneously blocking EGFR and COX-2

目的探讨厄罗替尼联合塞来昔布对肺腺癌A549细胞系凋亡、表皮生长因子受体(EGFR)和环氧合酶-2(COX-2)表达的影响。方法厄罗替尼、塞来昔布单独或联合干预细胞48 h后,倒置相差显微镜观察细胞形态;四甲基偶氮唑盐(MTT)法测定细胞抑制率;Hoechst33258法和TUNEL法检测细胞凋亡和流式细胞术检测细胞周期;免疫荧光检测EGFR和COX-2蛋白表达。结果厄罗替尼联合塞来昔布组相比单药组A549细胞明显出现大量颗粒和空泡,细胞变圆开始脱落;二者联合作用时抑制作用更强(P<0.05),厄罗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),并使细胞发生明显的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)。结论厄罗替尼与塞来昔布联合应用可协同介导细胞凋亡,阻滞细胞周期于G0/G1期及阻滞EGFR和COX-2信号途径。

Objective To explore the effects of erlotinib combined with celecoxib on apoptosis and the expression of epidermal growth factor receptor（EGFR） and cycloo xygenase-2（COX-2） in lung cancer A549 cells.MethodsA549 cells were observed under an inverted microscope,MTT was used to measure the growth inhibitory effects of A549 cells by erlotinib and celecoxib.The cell apoptosis was examined by TUNEL and Hoechst33258 staining method.The cell cycle was detected by flow cytometer,and the expression of EGFR,COX-2 were determined by immunofluorescence.Results A huge amount of granules and vacuolus were observed in the combined treatment A549,with cell rounding and defluxion（P〈0.05）.The combination increased apoptosis of A549（P〈0.05） and significantly induced G1 phase arrest（P〈0.05）and down-regulated expression of EGFR,COX-2 in A549 cells（P〈0.05）.Conclusion Combination therapy with erlotinib and celecoxib showed significant synergistic inducing apoptosis and G1 phase arrest,that is explained by down-regulating expressing both EGFR and COX-2.