NOX4抑制剂GKT137831增强顺铂对A549细胞的毒性作用

NOX4 inhibitor enhances the cytotoxicity of cisplatin in A549 cells

目的:研究GKT137831(NOX4抑制剂)和顺铂单用或联用对肺癌A549细胞增殖和凋亡的影响及作用机制.方法:MTT法检测不同浓度的GKT137831和顺铂对细胞毒性的影响,分别选取合适的浓度进行联合用药;流式细胞法检测细胞凋亡率;酶联免疫法检测Caspase3/7活性;免疫印迹检测A549细胞中Akt、p-Akt、MRP1蛋白表达.结果:GKT137831和顺铂单用均能抑制A549细胞的增殖,并诱导其凋亡,GKT137831可增强顺铂的细胞毒性.顺铂能上调p-Akt和MRP1的表达(P<0.05),GKT137831则抑制p-Akt和MRP1的表达(P<0.05);与对照组和单独用药组相比,联合用药能抑制p-Akt和MRP1的表达(P<0.05),表明GKT137831能抑制顺铂导致的p-Akt和MRP1表达上调.结论:GKT137831能够增强顺铂对肺癌细胞A549的毒性效应,作用机制可能通过抑制PI3K/AKT途径,调节其下游MRP1的表达有关.

Objective:To study the effect of GKT137831 and cisplatin alone or in combination on proliferation and apoptosis of lung cancer A549 cells.Methods:MTT assay was used to detect the effects of different concentrations of GKT137831 and cisplatin on cell survival,and the optimal concentration of each drug was selected for the following experiments.The apoptotic rate of cells was determined by flow cytometry.The activity of Caspase 3/7 was detected by enzyme-linked immunosorbent assay.Western blotting assay was used to measure the protein expression of Akt,p-Akt and MRP1 in A549 Cells.Results:GKT137831 or cisplatin alone inhibited proliferation and induce apoptosis of A549 cells.GKT137831 significantly enhanced cisplatin cytotoxicity.Cisplatin up-regulated expression of p-Akt and MRP1(P<0.05).GKT137831 inhibited expression of p-Akt and MRP1(P<0.05).Compared with the control group and the single drug group,the combined treatment group inhibited expression of p-Akt and MRP1(P<0.05),indicating that GKT137831 inhibited cisplatin-induced upregulation of p-Akt and MRP1.Conclusion:GKT137831 can strengthen the toxic effect of cisplatin on A549 cells,probably by inhibiting the PI3K/Akt pathway and regulating the expression of downstream MRP1.