白藜芦醇通过线粒体凋亡通路调节肺癌H460细胞增殖与凋亡的研究

摘要

目的:探讨白藜芦醇(Resveratrol,Res)对肺癌H460细胞增殖与凋亡的影响并探讨其可能的机制.方法:分别用浓度为0、25、100、200 μmol/L的Res作用于H460细胞48 h后,采用CCK-8法检测Res对H460细胞增殖的影响;流式细胞术检测细胞凋亡率;JC-1试剂盒检测细胞内线粒体膜电位水平;ATP试剂盒检测细胞内ATP水平;Western blot法检测细胞内Bcl-2、Bax和Cytochrome c蛋白表达情况.结果:与对照组比较,CCK-8及流式结果显示Res可抑制H460细胞的增殖,并使其凋亡率升高(P＜0.01);JC-1及ATP检测结果显示Res可降低H460细胞内线粒体膜电位及ATP水平(P＜0.01),且在该浓度范围内呈剂量相关性;Western blot检测发现随着Res浓度的增加,Bcl-2蛋白的表达水平下降,而Bax和Cytochrome c蛋白的表达水平升高.结论:实验浓度的Res可抑制H460细胞的增殖并诱导其凋亡,这一作用的机制可能与Res激活了线粒体凋亡通路进而诱发H460细胞凋亡有关.%Objective:To observe the effect of Resveratrol on proliferation and apoptosis of lung cancer H460 cells and explore the possible mechanisms.Methods:H460 cells were treated with Res (0,25,100,200 μmol/L) for 48 hours.The cell proliferation was measured by cell counting Kit-8,the apoptosis assays were performed by flow cytometry,the mitochondrial membrane potential level was detected by JC-1.The ATP levels in H460 cells were measured using an ATP assay kit,and the expression level of Bax,Bcl-2 and Cytochrome c protein in H460 cells were analysed by Western blot.Results:Compared with the control group,treated with Res inhibiteyd the proliferation of H460 cells,and enhanced the apoptosis rate through the CCK-8 and flow cytometry (P ＜0.01),and the JC-1 and ATP detection showed that Res decreased the mitochondrial membrane potential and ATP levels in H460 cells in a dose-dependent manner(P ＜ 0.01).Meanwhile,the expression level of Bcl-2 was downregulated,however the Bax and Cytochrome were upregulated with the increase of the Res concentration.Conclusion:Res inhibited the proliferation and induced the apoptosis of H460 cells,and which was involved in mitochondrial signaling pathway of apoptosis,potentially.