【摘要】 为了研究重组荞麦胰蛋白酶抑制剂(rBTI)诱导癌细胞凋亡的作用及其机制,采用细胞培养技术,以肝癌细胞系H22为靶细胞,以不同浓度的rBTI体外处理细胞不同时间后,采用MTT检测、DNA电泳分析、细胞核形态学观察、细胞色素c检测、caspases活性检测等方法测定rBTI在体外诱导肝癌细胞H22凋亡的作用。结果显示rBTI能够在体外特异性地抑制H22细胞的生长,诱导H22细胞凋亡,随着rBTI浓度及作用时间的增加,呈现剂量和时间依赖性效应。经rBTI处理的细胞可导致细胞色素c从线粒体释放到细胞质中,并能增强caspase-3、caspase-8、caspase-9的活性。结果表明,rBTI能特异性地诱导H22细胞凋亡,其机制与caspase-3依赖性凋亡调节信号通路有关。更多还原

【Abstract】 The aim of this study was to evaluate the in vitro anti-tumor effects of rBTI on hepatic cancer. Apoptosis of the H22 cell line induced by rBTI was identified by MTT assays, DNA electrophoresis analysis, morphological observation of nuclei, the measurement of cytochrome c and caspases activation assess. We found that rBTI inhibited cell viability by inducing apoptosis, as evidenced by the formation of apoptotic bodies, and DNA fragmentation. rBTI induced apoptosis was correlated with mitochondrial dysfunction, leading to the release of cytochrome c from the mitochondria to the cytosol, as well as the proteolytic activation of caspase-3, caspase-8 and caspase-9. Our results suggested that one of the pathways rBTI-induced apoptosis in H22 cells was mainly mediated by mitochondrial pathways via caspase-3. Moreover, rBTI could specifically inhibit the growth of cells of H22 hepatic carcinoma in vitro with a concentration-dependent and time-dependent effect, but there were minimal effects on normal mice liver cell.更多还原