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机构地区:[1]重庆医科大学病理生理学教研室干细胞与组织工程研究室,重庆400016
出 处:《中国生物制品学杂志》2010年第3期238-242,共5页Chinese Journal of Biologicals
摘 要:目的探讨鱼藤素诱导乳腺癌MDA-MB-231细胞凋亡的作用及其与线粒体凋亡途径的关系。方法用不同浓度的鱼藤素处理MDA-MB-231细胞,MTT法检测鱼藤素对MDA-MB-231细胞的增殖抑制作用;AnnexinV-FITC/PI双染流式细胞术检测细胞的凋亡率;罗丹明123单染法观察细胞线粒体膜电位的变化;Western blot法检测细胞cyt-c、caspase-3蛋白的表达水平;分光光度法检测细胞caspase-3蛋白活性。结果鱼藤素对MDA-MB-231细胞的增殖具有明显的抑制作用,且呈时效、量效依赖性;800和1200nmol/L鱼藤素处理组的细胞早期凋亡率显著高于未处理组;鱼藤素处理后,细胞线粒体膜电位降低,细胞浆内cyt-c和caspase-3蛋白的表达水平及caspase-3活性均明显升高。结论鱼藤素对乳腺癌MDA-MB-231细胞具有显著的增殖抑制、诱导凋亡作用,其作用与线粒体膜电位降低、释放细胞色素c,从而激活caspase-3依赖途径有关。Objective To investigate the effect of induction of breast cancer MDA-MB-231 cell apoptosis by deguelin as well as the relationship between the effect and chondriosome apoptosis pathway.Methods MDA-MB-231 cells were treated with deguelin at various concentrations.The inhibitory effect of deguelin on proliferation of MDA-MB-231 cells was determined by MTT method,the cell apoptosis rate by flow cytometry,the change of mitoehondrial membrane potential by Rhodamine 123 staining,the expression levels of cyt-c and caspase-3 by Western blot,and the caspase-3 protein activity by spectrophotometric method.Results Deguelin showed time-and dose-dependent inhibitory effect on proliferation of MDA-MB-231 cells.The apoptosis rates at early phase of MDAMB-231 cells treated with 800 and 1 200 nmol/L deguelin were significantly higher than those untreated.The mitoehondrial membrane potential of MDA-MB-231 cells after treatment with deguelin decreased,while the expression levels and activities of cyt-c and caspase-3 proteins increased significantly.Conclusion Deguelin inhibited the proliferation and induced the apoptosis of MDA-MB231 cells significantly,which were associated with the activation of caspase-3-dependent pathway by decreasing mitoehondrial membrane potential and releasing cyt-c.
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