二烯丙基三硫通过线粒体依赖性途径诱导人肝癌HepG2细胞凋亡  

Diallyl Trisulfide Induces Human Hepatocellular Carcinoma HepG2 Cell Apoptosis by Mitochondria-dependent Pathway

作　　者：肖志科[1] 张蒙夏[2] 姜浩[1] 伍尤华[1] 艾小红[1] 罗红梅[2] 汪煜华[2] 雷小勇[2] 唐圣松[2,1] 

机构地区：[1]南华大学附属第一医院肿瘤内科,衡阳421001 [2]南华大学药物药理研究所,药物蛋白质组学湖南省高等学校重点实验室,衡阳421001

出　　处：《生物化学与生物物理进展》2009年第10期1299-1305,共7页Progress In Biochemistry and Biophysics

基　　金：湖南省自然科学基金项目(08JJ5004,09JJ3060);湖南省科技计划重点项目(06FJ3203);湖南省教育厅科技计划重点项目(07A059)~~

摘　　要：二烯丙基三硫(diallyl trisulfide,DATS)对多种肿瘤有抗癌作用,但机制尚不完全清楚.为探讨DATS对人肝癌细胞系HepG2细胞凋亡的影响,用丫啶橙/溴化乙锭(AO/EB)法观察细胞凋亡情况,在显微镜下计数凋亡细胞数.JC-1荧光染色观察线粒体膜电势变化.Western blot法检测细胞色素c蛋白分布,ELISA法检测caspase-3活性.结果显示,DATS诱导HepG2细胞凋亡,用50μmol/L与100μmol/LDATS处理48h,细胞凋亡率分别达到60.33%和93.67%,并引起HepG2细胞线粒体膜电势降低.Western blot显示,DATS能诱导胞浆细胞色素c增加,与此同时,线粒体细胞色素c减少,诱导HepG2细胞caspase-3活化.提示:DATS可通过降低线粒体膜电势,促进细胞色素c由线粒体膜释放到胞浆中,激活caspase-3途径诱导人肝癌细胞系HepG2细胞凋亡.Diallyl trisulfide （DATS） is known to have an anticancer effect on human cancer cells. However, the exact mechanisms of this anticancer activity remain unclear. To explore the effect of DATS on human hepatocellular carcinoma HepG2 cell apoptosis. HepG2 cells were either treated with 50 μmol/L, 100μmol/L of DATS for 0, 6, 12, 24, 48 and 72 h, respectively. The mitochondrial membrane potential was visualized by the JC-1 fluorescence staining. The cytochrome c in HepG2 cells was detected by Western blotting. The activity of caspase-3 in HepG2 cells was measured with ELISA. Diallyl trisulfide induces HepG2 cell apoptosis. The apoptosis percentage in HepG2 cells is up to 60.33% and 93.67%, respectively, when the cell was treated with 50μmol/L or 100 μmol/L of DATS for 48 h. Diallyl trisulfide degrades the mitochondrial membrane potential and lowers the cytochrome c in mitochondria while increases the cytochrome c in the cytoplasma that mitochondria was deleted. Diallyl trisulfide activates caspase-3 kinase in HepG2 cells. These results indicate that diallyl trisulfide induces apoptosis by decreasing mitochondrial membrane potential, facilitating cytochrome c release from mitochondria into cytoplasm, and activating caspase-3 kinase in human hepatoceUular carcinoma HepG2 cell.

关 键 词：HEPG2细胞系 线粒体 二烯丙基三硫 细胞凋亡 

分 类 号：Q2[生物学—细胞生物学] R73[医药卫生—肿瘤]