摘要
目的:观察盐酸小檗碱对乳腺癌MDA-MB-231细胞抑制增殖及诱导凋亡的作用,并探讨其可能的机制。方法:不同浓度(0、25、50、75、100μmol/L)盐酸小檗碱处理MDA-MB-231细胞不同时间(48、72 h),MTT法检测盐酸小檗碱对细胞的增殖抑制率,选择盐酸小檗碱50μmol/L作为机制研究浓度,AnnenxinⅤ/PI EGFP染色流式细胞仪检测盐酸小檗碱对细胞凋亡的影响;2,7-二氯荧光黄双乙酸盐(DCFH-DA)荧光探针检测盐酸小檗碱对细胞内活性氧簇的影响;5,5′,6,6′-四氯-1,1′,3,3′-四乙基苯丙咪唑羰花青碘化物(JC-1)荧光探针检测盐酸小檗碱对细胞线粒体膜电位的影响;Caspase3试剂盒检测盐酸小檗碱对细胞内Caspase3活性的影响。结果:盐酸小檗碱作用MDA-MB-231细胞48、72 h,呈浓度及时间依赖性抑制MDA-MB-231细胞增殖。盐酸小檗碱(50μmol/L)呈时间依赖性诱导MDA-MB-231细胞凋亡并诱导MDA-MB-231细胞内活性氧簇大量生成,线粒体膜电位降低,Caspase3活性增高。结论:盐酸小檗碱对MDA-MB-231细胞的增殖抑制作用可能是通过诱导细胞内活性氧簇的生成、降低线粒体膜电位,升高Caspase3活性,由线粒体途径诱导细胞凋亡。
Objective：To investigate effects of berberine hydrochloride on breast cancer cells （MDA-MB-231） and related mechanism. Methods：Human breast cancer MDA-MB-231 cells were treated with different concentrations of berberine hydrochloride （0,10,25,50,75,100 μmol/L）,then the proliferation rate of MDA-MB-231 cells was examined by MTT assay after 48 h and 72 h;the apoptosis rate of MDA-MB-231 cells was detected by staning with Annenxin V/PI EGFP and flow cytometry;the level of reactive oxygen species in MDA-MB-231 cells was detected by DCFH-DA staining;the mitochondrial membrane potential was detected by molecular probe JC-1;activity of Caspase3 was detected by Caspase3 Kit. Results：Berberine hydrochloride inhibited the proliferation of MDA-MB-231 cells in a concentration-dependent manner. It also significantly elevated the reactive oxygen species level,reduced mitochondrial membrane potential and activity of Caspase3 in the MDA-MB-231 cells. Conclusion：Berberine can inhibit the proliferation and induce apoptosis of MDA-MB-231 cells through mitochondrial-dependent pathway.
出处
《南京医科大学学报（自然科学版）》
CAS
CSCD
北大核心
2013年第9期1220-1225,共6页
Journal of Nanjing Medical University(Natural Sciences)
基金
南京医科大学科技发展基金面上项目(09NJ-MUM099)
南京市医学科技发展青年启动项目(QYK10156)
关键词
乳腺癌
盐酸小檗碱
线粒体
活性氧簇
breast cancer
berberine hydrochloride
mitochondria
reactive oxygen species
