脂蟾毒配基诱导人肝癌细胞凋亡作用的研究

目的 通过体外脂蟾毒配基对人肝癌细胞（Bel7402）的凋亡诱导作用，为研究其抑制肿瘤细胞生长的作用机制提供依据。方法 通过应用流式细胞光度术检测细胞凋亡；采用细胞免疫细胞化学显色检测和Western blotting分析凋亡相关基因蛋白的表达来研究脂蟾毒配基对人肝癌细胞（Bel7402）凋亡的诱导作用。结果 表明脂蟾毒配基能够诱导Bel7402细胞发生凋亡，凋亡率大于50%；脂蟾毒配基提取液(浓度1.0 μm）作用于Bel7402细胞24小时后，bc1-2蛋白的表达下调，到48、72小时后下调更明显；而Bax蛋白的表达从24小时后开始上调，到48、72小时后表达上调明显。使用方差分析法与对照组相比较，P<0.05 ，统计学有显著意义。结论 提示诱导肿瘤细胞凋亡可能是脂蟾毒配基抑制、杀伤人肝癌细胞的机制之一。

The investigation of the effect on human Hepatoma Cell apoptosis induced by Resibufogenin

AIM to study the mechanism of Resibufogenin inhibited on the growth of human hepatoma cell by analyzing the apoptosis induced by Resibufogenin. METHODS The apoptosis was observed by means of Flow Cytometry. The effects of Resibufogenin on DNA synthesis and expression of correlative proteins in hepatoma cell apoptosis was observed by means of Western blotting and cytochemical method. RESULT It was showed that Resibufogenin could induce Bel7402 cell (human hepatoma cell) to apoptosis. The rate of apoptosis exceeds 50 percent. The expression of inhibiting apoptosis gene bcl-2 decrease after 24 hours in the group of 1.0 μm Resibufogenin and decreased more significantly after 48、72 hours; the expression of promoting apoptosis gene Bax increased after 24 hours and increased more significantly after 48、72 hours. CONCLUSION the resμlt indicate that inducing neoplasm cell apoptosis may be one mechanism of the Resibufogenin inhibiting and killing the human Hepatoma Cell .