Survivin在槲皮素诱导肝癌细胞SMMC7721凋亡中调节作用探究.docVIP

Survivin在槲皮素诱导肝癌细胞SMMC7721凋亡中调节作用探究 　【摘要】 目的研究槲皮素诱导肝癌细胞SMMC7721凋亡，探讨其诱导SMMC7721细胞凋亡与survivin蛋白表达和caspase-3活性的关系。方法采用MTT法检测槲皮素对肝癌SMMC7721细胞生长的抑制作用，Hoechst 33258荧光染色对细胞凋亡进行形态学检测，流式细胞术检测细胞周期变化，免疫细胞化学检测survivin蛋白表达，分光光度法检测caspase-3的活性变化。结果槲皮素抑制肝癌SMMC7721 细胞增殖的作用明显，且呈浓度和时间依赖性，槲皮素处理48 h的IC50为84.24 μmol·L-1。形态学检测显示出细胞凋亡的特征变化，流式细胞仪检测表明经槲皮素处理，肝癌SMMC7721 细胞周期阻滞于G0/G1期，且SMMC7721细胞survivin蛋白表达下降，caspase-3活性升高。结论槲皮素能诱导SMMC7721细胞凋亡，其机制可能是使肝癌SMMC7721 细胞周期阻滞于G0/G1期，并下调survivin蛋白的表达，直接激活caspase-3而诱导SMMC7721细胞凋亡。 【关键词】 槲皮素； 肝癌细胞SMMC7721； 细胞凋亡； Survivin； Caspase-3 Abstract：ObjectiveTo investigate the apoptosis of SMMC7721 hepatic cancer cells induced by quercetin and its relationship with the expression of survivin and activity of caspase-3.MethodsInhibitory effect of quercetin on SMMC7721 cells was detected by MTT assay. Morphological changes of apoptotic cells for Hoechst staining were observed by fluorescence microscope. Cell cycle was analyzed by flow cytometry (FCM). Expression of survivin protein was detected by immunocytochemistry staining. Activity of caspase-3 was analyzed by colorimetry. ResultsQuercetin had a significant inhibition on growth and proliferation of SMMC7721 cells in a concentration- and time-dependent manner. The IC50 was 84.24 μmol·L-1 after cells were treated with quercetin for 48h. evidence was provided that apoptosis occurred in SMMC7721 cells treated with quercetin using fluorescence microscope. It was suggested that quercetin blocked SMMC7721 cells at the G0/G1 phase by FCM analysis. Expression of survivin protein was decreased, and activity of caspase-3 were enhanced. ConclusionQuercetin could induce apoptosis of SMMC7721 cells by down-regcclating the expression of survivin protein and actionting caspase-3. Key words： Quercetin； Hepatic carcinoma cell lines SMMC7721； Apoptosis; Survivin； Caspase-3 槲皮素（Quercetin, Que, 3,5,7,3’,4’-五羟基黄酮）广泛存在于植物界，大约68 %的植物中都含有。槲皮素药理作用广泛，具有降血压、降血脂、扩张冠状动脉、抗血小板聚集、抗炎、抗过敏、抗心律失常和清除自由基等多种生物活性及药理作用［1～5］。近年发现槲皮素不仅对多种致癌剂、促癌剂有拮抗作