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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) has anticancer effect on many kinds of tumors, but the mechanism is not fully understood.In order to investigate the effect of DATS on the apoptosis of human hepatocellular carcinoma cell line HepG2, The apoptotic cells were observed by AO / EB method and the number of apoptotic cells was counted under the microscope.The change of mitochondrial membrane potential was observed by fluorescent staining of JC-1.Western blot was used to detect the distribution of cytochrome c protein and the expression of caspase-3 The results showed that DATS induced apoptosis in HepG2 cells, and the apoptotic rates of HepG2 cells treated with 50μmol / L and 100μmol / L LDAT for 48h were 60.33% and 93.67%, respectively, and the mitochondrial membrane potential of HepG2 cells was decreased.Western blot showed that DATS Induced the increase of cytochrome c in the cytoplasm, meanwhile mitochondrial cytochrome c decreased and induced the activation of caspase-3 in HepG2 cells.Conclusion: DATS can release mitochondrial membrane potential and promote the release of cytochrome c from the mitochondrial membrane into the cytoplasm and activate caspase-3 pathway induces apoptosis in human hepatoma cell line HepG2.