白藜芦醇抑制人结肠癌细胞增殖及其与PTEN的关系研究.docVIP

DOI：10.16016/j.1000-5404.201410125 白藜芦醇抑制人结肠癌细胞增殖及其与PTEN的关系研究 胡 培，伍秋香，袁霜雪，王东旭，刘映孜，何百成，孙文娟 （400016重庆，重庆医科大学药学院药理教研室，重庆市生物化学与分子药理学重点实验室） [摘要] 目的 研究白藜芦醇（resveratrol, Res）对人结肠癌细胞增殖的抑制作用及可能的机制。方法 采用结晶紫染色、流式细胞术和Western blot分析Res对HCT116细胞增殖的抑制作用；应用流式细胞术和免疫荧光分析Res对HCT116细胞凋亡的促进作用；采用Western blot和PCR分析Res对HCT116细胞内Akt1/2及PTEN表达水平的影响；通过重组腺病毒介导的PTEN过表达或沉黙表达，分析Res抑制HCT116细胞增殖及其与PTEN的关系。结果 与对照组相比，Res处理组HCT116细胞增殖受到抑制（P0.05, P0.01），G1期细胞比例也随Res浓度增加而升高，同时PCNA蛋白表达水平下降；流式细胞术分析结果显示，Res 能明显促进HCT116细胞凋亡; Western blot和PCR分析结果显示，Res对Akt1/2 mRNA表达无明显影响，但能降低Akt1/2的磷酸化水平，同时促进PTEN表达。外源性过表达PTEN能增强Res对HCT116细胞的增殖抑制作用(P0.01)，而沉黙PTEN表达能部分逆转Res对HCT116细胞增殖的抑制作用(P0.01)。结论 Res对HCT116细胞增殖具有抑制作用，并能促进其凋亡；Res的这种作用可能与其促进PTEN表达，抑制PI3K/Akt活化有关。 [关键词] 白藜芦醇；结肠癌；增殖抑制；凋亡；PTEN；PI3K/Akt [中图法分类号] [文献标志码] A Relationship between proliferation inhibition effect of resveratrol and PTEN in colon cancer cells Hu Pei, Wu Qiuxiang, Yuan Shuangxue, Wang Dongxu, Liu Yingzi, He Baicheng, Sun Wenjuan (Chongqing Key Laboratory of Biochemistry and Molecular Pharmacology, Department of Pharmacology, Chongqing Medical University, Chongqing, 400016, China) [Abstract] Objective To investigate the effect of resveratrol (Res) on inhibiting proliferation of colon cancer cells and explore the possible mechanism. Methods The proliferation inhibition effect of Res in HCT116 colon cancer cells was measured by crystal violet staining, flow cytometry and Western blot. Then, the apoptosis was detected by immunofluorescence assay and flow cytometry. The expressions of Akt1/2 and PTEN and the phosphorylation of Akt1/2 in HCT116 cells were analyzed by RT-PCR and Western blot. Finally, the effect of PTEN on the proliferation inhibition effect of Res was evaluated by recombinant adenovirus for PTEN over-expression or knockdown in HCT116 cells. Results Res inhibited the proliferation of HCT116 cells in a concentration-dependent manner (P0.05 and P0.01), arrested the cell cycle at G1 phase,