摘要
目的:探讨去甲斑蝥素(NCTD)对人肝癌SMMC-7721细胞凋亡的影响及其机制。方法:分别用MTT和AnnexinⅤ/PI法检测NCTD对人肝癌细胞SMMC-7721的生长抑制率和细胞凋亡率。用流式细胞仪检测线粒体膜电位的变化。应用Western blot方法来分析细胞色素c、caspase-3、AIF、Bcl-2和Bax的表达。结果:NCTD对人肝癌细胞SMMC-7721有生长抑制作用,随浓度升高、时间延长作用增强,呈剂量和时间效应关系。用AnnexinⅤ/PI双染色法检测SMMC-7721细胞,随NCTD的剂量增加,细胞凋亡率也增加,呈剂量依赖性。用流式细胞仪检测细胞膜电位,随NCTD的剂量增加,线粒体膜电位也随之下降。应用Western blot analysis方法检测,NCTD可诱导caspase-3和AIF的活化及线粒体细胞色素c释放到胞浆中。并随NCTD的剂量增加,Bax的表达量上升,而Bcl-2的表达量下降。结论:NCTD能显著抑制SMMC-7721细胞增殖,并可通过内源性线粒体信号转导途径诱导细胞凋亡。
Objective:To investigate the mechanism of norcantharidin (NCTD)-induced apoptosis of SMMC-7721 hepatoma cells.Methods:The growth inhibition of SMMC-7721 cells was measured by MTT method.Apoptosis was detected by Annexin Ⅴ/propidium iodide staining.The mitochondrial membrane potential was measured by flow Cytometry.Western blot analysis was used to evaluate the expressing level of Cyeochrome c,caspase-3,AIF,Bcl-2 and Bax.Results:Norcantharidin inhibited SMMC-7721 cell growth in a time-and dose-dependent manners.SMMC-7721 cells treated with norcantharidin were showed by typical characteristics of apoptosis.The cells treated with NCTD were showed the loss of mitochondrial membrane potential.The activities of caspase-3,Cyeochrome c,AIF,and Bax were up-regulated after NCTD treatment at different doses.The expression of Bcl-2 was decreased after treatment with NCTD.Conclusion:These results suggest that NCTD induces apoptosis in SMMC-7721 cells via activation of the mitochondrial pathways.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2010年第7期611-615,共5页
Chinese Journal of Immunology