白藜芦醇诱导鼻咽癌CNE-1细胞凋亡及其分子机制

Studied on Resveratrol in Inducing apoptosis of Nasopharyngeal Carcinoma CNE-1 Cells and Its Molecular Mechanism

目的探讨白藜芦醇诱导鼻咽癌CNE-1细胞凋亡及其分子机制。方法噻唑蓝(MTT)比色法检测细胞活力;活细胞染色法(PI)流式细胞仪分析细胞周期和PI-AnnexinV双染色法流式细胞仪检测细胞凋亡;免疫印迹检测蛋白表达水平。结果 MTT结果表明白藜芦醇呈浓度和时间依赖性抑制鼻咽癌CNE-1细胞活力,与正常对照组比较,差异有统计学意义(P<0.01);PI单染流式细胞仪分析表明白藜芦醇引起S期的CNE-1比例明显增多,相应免疫印迹结果显示细胞周期蛋白Cyclin E和Cyclin A显著下调,呈药物浓度依赖;PI-Annexin V双染流式细胞仪分析表明白藜芦醇呈药物浓度依赖诱导CNE-1细胞凋亡;免疫印迹分析显示抗凋亡蛋白Bcl-2表达下调,而促凋亡蛋白Bax表达上调。结论白藜芦醇能抑制鼻咽癌CNE-1细胞的增殖,使其阻滞在S期,并可能通过线粒体介导的凋亡通路诱导其凋亡。

Objective To elucidate the effects of resveratrol on inducing apoptosis of nasopharyngeal carcinoma CNE-1 cells and its molecular mechanism. Methods The viability of CNE-1 cells was detected by methyl thiazolyl tetrazolium（MTT） assay, cell cycle profile was analyzed by PI single-staining flow cytometer and apoptosis of CNE-1 cells was analyzed by PI-AnnexinV double-staining flow cytometer. The expression level of target proteins was examined by immunoblotting method. Results Resveratrol significantly inhibited the viability of CNE-1 cells inconcentration- and time-dependent manner （P 〈 0.01 compared with the normal control group）. Treatment with resveratrol resulted in the increased Cyclin E and Cyclin A were the percentage of S-phase CNE-1 cells in dose-dependent manner. Accordingly, downregulated in resveratrol-treated CNE-1 cells. Furthermore, resveratrol dose-dependently induced the apoptosis of CNE-1 cells. Correspondingly, the anti-apoptotic protein Bcl-2 was downregulated and the pro-apoptotic protein Bax was upregulated in CNE-1 cells after treatment with resveratrol. Conclusion Resveratrol can arrest CNE-1 ceils at S phase via downregulating the expression of cyclinE and Cyclin A, and can further induce apoptosis through mitochondrial-mediated intrinsic cell-death pathways.