盐酸小檗碱抑制乳腺癌MDA-MB-231细胞增殖及诱导凋亡的作用研究
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南京医科大学科技发展基金面上项目(09NJMUM099);南京市医学科技发展青年启动项目(QYK10156)


Effects of berberine hydrochloride on breast cancer cells (MDA-MB-231)
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    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(0-25-50-75-100 μmol/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50 μmol/L作为机制研究浓度,Annenxin Ⅴ/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(50 μmol/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。

    Abstract:

    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.

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谢 娟,黄新艳,许银燕,陈燕妮,汤 倩.盐酸小檗碱抑制乳腺癌MDA-MB-231细胞增殖及诱导凋亡的作用研究[J].南京医科大学学报(自然科学版),2013,(9):1220-1225

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  • 收稿日期:2013-04-09
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  • 在线发布日期: 2013-09-18
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