构树叶总黄酮提取物诱导人肝癌细胞HepG2细胞凋亡及机制  

Study on the effects of TFBP on apoptosis of human hepatoma carcinoma cell line HepG2

作　　者：朱开梅[1] 李美波[1] 骆彩珍[2] 刘建楠[1] 顾生玖[1] 

机构地区：[1]桂林医学院药学院,广西桂林541004 [2]桂林医学院护理学院,广西桂林541004

出　　处：《中国医院药学杂志》2013年第21期1763-1767,共5页Chinese Journal of Hospital Pharmacy

基　　金：广西科技特派员专项资助项目(编号:桂科能129825-21);广西教育厅项目资助项目(编号:201202ZD065);桂林市科技攻关资助(编号:20110106-5;20120105-5;20120105-16;20120105-8)

摘　　要：目的:研究构树叶总黄酮(total flavonoids of Broussonetia papyrifera,TFBP)对人肝癌细胞HepG2的生长抑制和诱导凋亡的作用及其机制。方法:采用MTT法检测细胞生长抑制率;Hoechst 33342荧光染色法荧光显微镜观察细胞的形态变化;流式细胞仪检测细胞的凋亡率;RT-PCR法检测基因Bcl-2、Bax mRNA的表达;免疫细胞化学SP法检测基因Bcl-2、Bax蛋白表达。结果:MTT结果显示,TFBP对HepG2有增殖抑制作用,并存在浓度和时间依赖关系;Hoechst 33342荧光染色可观察到核浓缩及核碎裂等细胞凋亡特征;流式细胞仪结果显示,细胞出现凋亡形态改变,且细胞被阻滞于G2/M期,随浓度增加,凋亡率逐渐增加;RT-PCR和SP结果显示,随着TFBP浓度增加Bcl-2 mRNA和蛋白表达逐渐减弱,Bax mRNA和蛋白的表达逐渐增强。结论:TFBP在体外对肝癌细胞HepG2有明显的增殖抑制和诱导细胞凋亡的作用。其诱导凋亡的机制可能与其下调Bcl-2蛋白和上调Bax蛋白的表达有关。OBJECTIVE To investigate the effects of total flavonoids of Broussonetia papyrifera（TFBP） on antitumor activity and induction of apoptosis in vitro, using human hepatoma carcinoma cell line HepG2. METHODS The viability of HepG2 cells was measured by MTT assay. Morphology of cell apoptosis was observed by Hoechst 33342 fluorescence staining. The percentage of apoptotic cell was detected by flow cytometry. The gene expression of Bax and Bcl-2 was detected hy RT-PCR. The protein expression of Bcl-2 and Bax was analyzed by immunocytochemical method（SP）. RESULTS TFBP inhibited the growth of cells and caused apoptosis significantly. The suppression was both in a time-and dose-dependent manner, the ratio of FCM G2/M cells increased and S cells decreased, at the same time, TFBP down-regulated the Bcl 2 expression and up-regula- ted the Bax expression. CONCLUSION TFBP has apparent inhibitory action and apoptosis-inducing effect on HepG2 cells. TFBP may induce apoptosis via down-regulating Bcl-2 and up regulating of Bax in HepG2 cells.

关 键 词：HEPG2细胞株 构树叶总黄酮 细胞凋亡 BCL-2 BAX 

分 类 号：R285.5[医药卫生—中药学]