Salinomycin对乳腺癌阿霉素耐药细胞株MCF-7/DOX的增殖抑制作用及机制  

Anti-proliferative effect of salinomycin on doxorubicin- resistant human breast cancer MCF-7/DOX cells

作　　者：刘浩[1] 卢敏莹[1] 贺智敏[1] 

机构地区：[1]广州医科大学附属肿瘤医院肿瘤研究所,广东广州510095

出　　处：《中国药理学通报》2015年第4期549-554,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81402497)

摘　　要：目的本研究旨在探讨Salinomycin对乳腺癌阿霉素耐药细胞株MCF-7/DOX增殖和凋亡的影响及可能作用机制。方法 MTS实验检测Salinomycin对MCF-7/DOX细胞增殖的影响;Annexin V-FITC/PI染色检测Salinomycin对MCF-7/DOX耐药细胞凋亡的影响;DCFH-DA染色检测Salinomycin对MCF-7/DOX耐药细胞活性氧(reactive oxygen species,ROS)产生的影响;JC-1法测定细胞线粒体膜电位;Western blot法检测细胞凋亡相关蛋白BAX、BCL-2、caspase-3和caspase-9的表达变化。结果 Salinomycin能明显抑制MCF-7/DOX耐药细胞增殖,且具有浓度依赖性;流式分析发现Salinomycin能够诱导MCF-7/DOX细胞凋亡,增加细胞内ROS水平,降低细胞线粒体膜电位;与对照组相比较,Salinomycin处理明显抑制BCL-2的表达,上调BAX、cleaved caspase-3和cleaved caspase-9的蛋白表达;抗氧化剂N-acetylcysteine(NAC)则逆转上述作用。结论 Salinomycin能够诱导MCF-7/DOX细胞凋亡,其机制可能与Salinomycin诱导ROS的产生,激活线粒体凋亡途径有关。Aim To investigate the anti-proliferative effect of salinomycin on doxorubicin-resistant human breast cancer MCF-7 / DOX cells. Methods MCF-7and MCF-7 / DOX cells were treated or untreated with salinomycin. Cell viability was detected by MTS assay.Cell apoptosis was detected by Annexin V-FITC / PI assay. Reactive oxygen species（ ROS） was measured by DCFH-DA staining. Mitochondrial membrane potential was measured by JC-1 assay. The expression of apoptosis related proteins BAX, BCL-2, caspase-3, and caspase-9 were evaluated by Western blot analysis.Results The cell viability was significantly reduced by salinomycin treatment in a dose-dependent manner.The flow cytometry results showed that salinomycin induced MCF-7 / DOX cell apoptosis,increased ROS production,and decreased mitochondrial membrane potential. Furthermore,salinomycin decreased the expression of BCL-2,and increased the expression of BAX,cleaved caspase-3,and cleaved caspase-9. Moreover,the antioxidant N-acetylcysteine（ NAC） markedly blocked the above effects. Conclusions Our results suggest that salinomycin-induced apoptosis in MCF-7 /DOX is associated with induction of ROS production,and activation of mitochondria apoptosis pathway,which may become a potential chemotherapeutic agent for the therapy of doxorubicin resistant breast cancer.

关 键 词：乳腺癌 SALINOMYCIN 阿霉素耐药 细胞凋亡 活性氧 线粒体膜电位 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学]