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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,MTT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-2mRNA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 and mechanism of bufalin on proliferation and apoptosis of human breast cancer cell line MDA-MB-231 in vitro. Methods MDA-MB-231 cells were treated with different concentrations of bufalin. Cell viability was measured by MTT assay. Apoptotic cells were detected by flow cytometry. The levels of reactive oxygen species ROS), and the expression of Bax and Bcl-2 mRNA were detected by RT-PCR. Results Bufalin could decrease the survival rate of MDA-MB-231 cells and induce apoptosis in a dose-dependent manner (P <0.01). After treatment with bufalin, intracellular ROS level increased (P <0.01), Bax mRNA increased, and Bcl-2 mRNA decreased (P <0.05 or P <0.01). Conclusions Bufalin can inhibit the proliferation and induce the apoptosis of human breast cancer MDA-MB-231 cells in vitro, which may be related to the increase of intracellular ROS level and the up-regulation of Bax / Bcl-2 ratio.