白藜芦醇诱导肺癌细胞凋亡和细胞周期阻滞的分子机制

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。

Molecular mechanism of resveratrol inhibiting proliferation of human lung cancer cells through inducing cell cycle arrest and apoptosis

Objective To elucidate the potential action mechanisms of resveratrol on human lung cancer cell lines.Methods The MTT method was used to detect the cell proliferation activity of the lung cancer cell lines A549 and H1299, and the Western blot method was used to detect the expressions of the apoptosis related proteins (cleaved caspase-3, cleaved caspase-7, cleaved caspase-9, and cleaved PARP) and the cell cycle related proteins (cyclin D1, cyclin D3, CDK2, CDK4, CDK6, p53, p18, p21 and p27).The flow cytometry was used to detect the cell cycle change and apoptosis situation of resveratrol-treated cell populations.Results Resveratro] displayed the concentration-dependent inhibitory effect in lung cancer cell lines A549 and H1299 growth.Treated with resveratrol, the expression levels of cyclin D1, cyclin D3 and CDK2 proteins were up-regulated but the expression levels of CDK4, CDK6, p53, p18, p21 and p27 were down-regulated in A549 and H1299 cell lines.There was an obvious up-regulation to the expression levels of cleaved caspase-3, cleaved caspase-7, cleaved caspase-9, cleaved PARP in A549 and H1299 cell lines with resveratrol treatment.The flow cytometry detection showed that with the increase of the concentration of resveratrol, the apoptosis proportion increased in A549 and H 1299 lung cancer cell lines and the cell cycle was blocked in G1/S phase obviously.Conclusions Resveratrol can promote the caspase activation and cell apoptosis, and down-regulate the expression levels of cyclin D1, cyclin D3 and CDK2, but up-regulate the expression levels of CDK4, CDK6, p53, p18, p21 and p27, which may be the key mechanisms that resveratrol induces G1/S phase block in human lung cancer cells.