盐酸小檗碱对人胃癌细胞增殖和凋亡的影响

摘要

Objective : : To investigate the effect of Berberine on the proliferation and apoptosis in two gastric cancer cell lines and one gastric epithelial cell line GES, and to elucidate the underlying mechanism. Methods : MTT assay was adopted to detect the proliferation state of cells, and FCM was performed to found the alterations of cell cycle and apoptosis. Furthermore , we examined the expression of proteins which were associated with cell cycle and apoptosis by Western blotting. Results :The growth of gastric cancer cells were inhibited by Berberine( P ＜ 0. 05 ) ; Berberine induced a dose - dependent inhibition of cell growth in all three cell lines, and 50μg/ml of Berberine induced 81. 3％ growth inhihition in gastric cancer cells. As shown by flow cytometry, a low concentration of Berberine ( lOμg/ml ) could cause G0/G1 cell cycle arrest and cell apoptosis( P ＜ 0. 05 ). The result of Western blot indicated that Berberine suppressed the protein expression of Bcl - 2, and up - regulate the protein expression of Bax and p53 in gastric cancer cells in a time - and dose - dependent manner. Conclusion: Berberine inhibits the growth of gastric cancer cells by inducing cell cycle arrest and apoptosis. The underlying mechanism was partly due to the down regulaion of Bcl -2 and the up - regulation of Bax and p53 in a time - and dose - dependent manner.%目的:研究盐酸小檗碱对人胃癌细胞及永生化胃上皮细胞增殖和凋亡的影响,初步探讨其中可能的分子机制.方法:利用不同浓度的小檗碱处理胃癌细胞系MKN-45和SGC-7901细胞,以及永生化胃上皮细胞系GES细胞,采用MTT法测定细胞活力,流式细胞术检测细胞周期和细胞凋亡(PI标记),Western blot检测细胞周期和凋亡相关蛋白的表达.结果:MTT结果显示与空白对照组相比,小檗碱处理后,细胞增殖速度明显受到抑制(P<0.05),并呈浓度依赖性,其中50μg/ml处理组最大抑制率为81.3%,这种抑制作用对胃癌细胞更为明显.流式细胞仪检测表明,小檗碱在较低浓度(10μg/ml)时即可诱导胃癌细胞出现G0/G1阻滞,S期细胞百分比减少和细胞凋亡(P<0.05),这种作用呈时间和浓度依赖性.Western blot结果提示小檗碱处理后,胃癌细胞Bcl-2蛋白表达水平降低,Bax及p53蛋白表达水平明显上调.结论:小檗碱可抑制胃癌细胞增殖,诱导胃癌细胞凋亡,其机制与阻滞细胞周期进展以及调控凋亡相关蛋白Bcl-2、Bax和p53密切相关.