二烯丙基二硫通过激活Caspase3和释放Cyt-c诱导Raji细胞凋亡

DIALLYL DISULFIDE-INDUCED APOPTOSIS OF RAJI CELLS IS MEDIATED BY CASPASE ACTIVATION AND CYTOCHROME C RELEASE

二烯丙基二硫(diallyl disulfide,DADS)作为天然植物大蒜中的提取物,能抑制多种肿瘤细胞生长,但其抑瘤的分子机制还不十分清楚。在该研究中,作者采用CCK-8(cell counting ki)t技术检测发现,DADS能有效地抑制人淋巴瘤Raji细胞增殖,形态学观察、DNA琼脂糖凝胶电泳和流式细胞仪检测证实DADS呈时间和浓度依赖性诱导Raji细胞凋亡,DADS处理细胞24h后,MCL1和Bcl-2蛋白表达下降,而Bax和Bak蛋白表达水平无变化,Bid和Caspase3被激活,线粒体中Cyt-c释放增多,用Caspase抑制剂Z-VAD-FMK能部分阻断DADS诱导人淋巴瘤Raji细胞凋亡,提示DADS诱导的Raji细胞凋亡作用通过Bcl-2/MCL1-线粒体-caspase3通路介导。

Diallyl disulfide （DADS） is a natural organosulfur compound derived from crushed garlic, it has been reported that the antiproliferative activity of DADS in several cancer cell lines. However, the molecular mechanism underlying the DADS-antitumor effect is not yet clearly understood. In the present study, the CCK-8 was used by assay the inhibition of DADS on the cells viability, the DADS-induced apoptosis in human Raji cells were investigated using both morphological analysis and DNA fragmentation technique. In this study, demonstrated that DADS-induced apoptosis in human Raji cells is a concentration- and time-dependent manner. This DADS-induced apoptosis in human Raji cells was also accompanied by the down-regulation of Bcl-2 and MCL-1 and activation of Bid, but no effect on the levels of Bax and Bak. Moreover, the DADS treatment resulted in increase in the release of mitochondrial cytochrome c （cyt c）, and activation of caspase 3. Furthermore, treatment with caspase inhibitor （z-DEVD-fmk） was capable of preventing the DADS-induced caspase 3 activity and cell death. These results clearly demonstrate that the induction of apoptosis by DADS involves be mediated, at least in part, by the Bcl-2/MCLl-mitochondria-Caspase3 pathway in human Raji cells.