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目的探讨白藜芦醇对肺癌细胞系A549和H1299细胞增殖与细胞周期阻滞及细胞凋亡之间的联系。方法采用MTT法检测白藜芦醇对A549和H1299细胞增殖的影响;用Western印迹法检测白藜芦醇作用后A549和H1299中凋亡相关蛋白cleaved-caspase-3、cleaved-caspase-7、cleaved-caspase-9、cleaved-PARP及细胞周期相关蛋白cyclin D1、cyclin D3和CDK2、CDK4、CDK6、p53、p18、p21、p27的表达情况;通过流式细胞仪技术检测白藜芦醇诱导A549和H1299细胞凋亡率以及细胞周期阻滞的影响。结果白藜芦醇对A549和H1299的生长均有抑制作用,呈浓度依赖性;经白藜芦醇处理后,A549和H1299的周期蛋白cyclin D1、cyclin D3和CDK2蛋白的表达水平上调,而周期蛋白CDK4、CDK6、p53、p18、p21、p27的表达水平下调;A549和H1299的凋亡相关蛋白cleaved-caspase-3、cleaved-caspase-7、cleaved-caspase-9、cleaved-PARP的表达水平上调;流式细胞仪检测A549和H1299的细胞凋亡率随白藜芦醇的浓度上升而增加,流式细胞仪检测细胞周期即可观察到随白藜芦醇的浓度的增加,细胞周期明显阻滞于G1/S期。结论白藜芦醇可能是通过促进caspases通路的活化,从而促进细胞凋亡;周期蛋白cyclin D1、cyclin D3和CDK2蛋白的表达水平的上调和CDK4、CDK6、p53、p18、p21、p27的表达水平的下调可能是白藜芦醇诱导人肺癌细胞发生G1/S期阻滞的关键机制。
Objective To investigate the relationship between resveratrol and lung cancer cell lines A549 and H1299 cell proliferation and cell cycle arrest and apoptosis. Methods The effects of resveratrol on the proliferation of A549 and H1299 cells were detected by MTT assay. The apoptosis-related proteins cleaved-caspase-3, cleaved-caspase-7 and cleaved were detected by Western blotting in A549 and H1299 -caspase-9, cleaved-PARP and cyclin D1, cyclin D3 and CDK2, CDK4, CDK6, p53, p18, p21 and p27 were detected by flow cytometry.Results The resveratrol- H1299 cell apoptosis rate and cell cycle arrest. Results Resveratrol could inhibit the growth of A549 and H1299 in a concentration-dependent manner. After resveratrol treatment, the expressions of cyclin D1, cyclin D3 and CDK2 were up-regulated in A549 and H1299 cells, The expressions of cleaved-caspase-3, cleaved-caspase-7, cleaved-caspase-9 and cleaved-PARP in A549 and H1299 were up-regulated ; Flow cytometry A549 and H1299 apoptosis rate increased with the increase of resveratrol, flow cytometry cell cycle can be observed with the concentration of resveratrol increased significantly cell cycle resistance Lag in G1 / S period. Conclusion Resveratrol may promote the apoptosis through promoting the activation of caspases pathway. The expressions of cyclin D1, cyclin D3 and CDK2 protein and the expressions of CDK4, CDK6, p53, p18, p21, p27 Down-regulation may be the key mechanism of resveratrol-induced G1 / S arrest in human lung cancer cells.