摘要
目的探讨三羟异黄酮(genistein)对人肝癌细胞凋亡的诱导作用及其机制。方法将三羟异黄酮作用于体外培养的人肝癌细胞(SMMC-7721),MTT法检测增殖抑制率,光镜下观察凋亡细胞的形态,凝胶电泳分析DNA改变;通过免疫组化技术检测凋亡基因蛋白P53、survivin和caspase-3的表达。结果三羟异黄酮呈浓度和时间依赖性抑制人肝癌细胞的增殖。5mg/L、10mg/L和20mg/L三羟异黄酮作用48h对肝癌细胞的抑制率分别为9·86%、19·13%和25·64%。形态学显示,三羟异黄酮能诱导SMMC-7721细胞凋亡,细胞DNA电泳出现典型梯状条带;且凋亡相关基因蛋白survivin表达明显减少,而P53和caspase-3的表达增加。结论三羟异黄酮能诱导SMMC-7721细胞凋亡,其凋亡机制之一可能是下调survivin基因蛋白表达,增强P53和caspase-3的表达。
Objective To study the apoptosis of human hepatocarcinoma cells induced by genistein and its mechanism. Methods Human hepatocarcinoma SMMC-7721 cells cultured in vitro were treated with genistein. MTr test was used to examine the proliferation inhibitory rate, The morphology of apoptotic cells was observed by microsco- py. DNA fragmentation was examined by gel electrophoresis. The expressions of apoptotic gene of P53,SURVIVIN and caspase-3 were examined by immunohistochemstry method. Results Genistein inhibited proliferation of human hepatoma cells in a dose and time-dependent manner, The inhibitory rates were 9.86% ,19. 13% and 25.64% in 5,10 and 20 mg/L for 48 h. It also induced apoptosis of SMMC-7721 cells, DNA of cells presented ＂ladder＂ break. The expression of apoptosis related protein survivin was down regulated, the expressions of P53 and caspase- 3 were up regulated. Conclusion Genistein can induce apoptosis of SMMC-7721 cells. One of the molecular mechanisms of genistein might be inhibition of apoptosis related protein expression of survivin and increase of expressions of P53 and caspase-3.
出处
《基础医学与临床》
CSCD
北大核心
2007年第8期881-885,共5页
Basic and Clinical Medicine
基金
河北省卫生厅资助项目(04009)
