蟾毒灵对人乳腺癌MDA-MB-231细胞增殖和凋亡的作用

摘要

目的 探讨蟾毒灵在体外对人乳腺癌MDA-MB-231细胞增殖和凋亡的作用及初步机制.方法 以不同浓度的蟾毒灵处理MDA-MB-231细胞,M T T 法检测细胞存活率,流式细胞术检测细胞凋亡水平,2',7'-二氯荧光黄双乙酸盐法测定活性氧(ROS)水平,RT-PCR检测细胞凋亡相关基因Bax和Bcl-2 mRNA表达水平.结果 蟾毒灵可剂量依赖性地降低MDA-MB-231细胞的存活率,诱导细胞凋亡(P<0.01).蟾毒灵处理细胞后,细胞内ROS水平增加(P<0.01),Bax mRNA表达增加,而Bcl-2 mRNA 表达减少(P<0.05或P<0.01).结论 蟾毒灵在体外能抑制人乳腺癌MDA-MB-231细胞的增殖,诱导细胞凋亡,该作用可能与其升高细胞内ROS水平、上调Bax/Bcl-2的比例有关.%Objective To investigate the effect of bufalin on human breast cancer MDA-MB-231 cells proliferation and apoptosis in vitro and its related molecular mechanism.Methods After MDA-MB-231 cells were cultured and treated with different concentrations of bufalin,the cell survival rate was measured by MTT,the cell apoptosis was analyzed by flow cytometry,the content of reactive oxygen species(ROS)was examined by 2',7'-dichlorofluorescein diacetate assay,and the mRNA expressions of Bax and Bcl-2 were detected by RT-PCR,respectively.Results With bufalin treatment,the survival rate of MDA-MB-231 cells was decreased in a dose-dependent manner and the apoptosis was induced (P<0.01).Bufalin could increase the level of ROS(P<0.01),and upregulate the expression of Bax and downregulate the expression of Bcl-2 at the mRNA level(P<0.05 or P<0.01).Conclusion Bufalin is able to inhibit the proliferation and induce apoptosis of MDA-MB-231 cells in vitro,and the possible mechanism may be related with the effect of increasing ROS level and upregulating the ratio of Bax/Bcl-2.