过表达HIF-2α对肝癌细胞运动和侵袭的影响  

Effect of overexpression of HIF-2 alpha on movement and invasion of hepatocellular carcinoma cells

作　　者：孙海香[1] 杨柳晓[1] 

机构地区：[1]复旦大学附属中山医院肝癌研究所,上海200032

出　　处：《中国临床医学》2016年第3期334-336,共3页Chinese Journal of Clinical Medicine

基　　金：国家自然科学基金(81302100;81502028;81572884d;81372317);高等学校博士学科点专项科研基金(20120071120068);复旦大学附属中山医院优秀青年基金(2015ZSYXQN03)~~

摘　　要：目的:探讨缺氧诱导因子(hypoxia inducible factor-2α,HIF-2α)对肝癌细胞运动和侵袭的影响。方法:构建HIF-2α过表达载体,转染肝癌细胞,构建过表达稳定表达细胞株。采用实时荧光定量多聚酶联反应(quantitative real-time polymerase chain reaction,qRT-PCR)和蛋白质印迹法检测HIF-2α表达水平,通过Transwell运动实验、划痕实验体外分析过表达HIF-2α对肝癌细胞运动的影响;将过表达HIF-2α细胞原位接种裸鼠,观察小鼠肺脏的转移情况。采用蛋白质印迹法检测EMT相关指标分析HIF-2α促进肝癌细胞运动、侵袭能力的作用机制。结果:将过表达载体转染肝癌细胞后,HIF-2αmRNA和蛋白表达水平均升高(P<0.05)。Transwell运动实验发现,高表达HIF-2α后,穿膜细胞数目明显增加(P<0.01);划痕实验结果也表明高表达HIF-2α的细胞运动能力增强。体内侵袭实验发现高表达HIF-2α组小鼠肺转移灶大小增加。蛋白质印迹结果显示HIF-2α可促进间质细胞相关标志物蛋白表达增加。结论:过表达HIF-2α可能通过促进肝癌细胞的EMT转化提高肝癌细胞的运动、侵袭能力。Objective：To investigate the effects of hypoxia inducible factor 2 alpha（HIF-2α）on movement and invasion of hepatocellular carcinoma cells（HCC）.Methods：The plasmid of over-expression of HIF-2α was constructed and transfected into HCC cells,then the stable expression cell strains were selected.HIF-2α expression levels were detected by quantitative real-time polymerase chain reaction（RT-PCR）and Western Bloting.The effects of HIF-2α over-expression on the movement ability of HCC were analyzed in vitro by Transwell assay and scrach assay.The over-expressed HIF-2α cells were planted in situ in nude mice,and the lung metastasis were observed.we checked the EMT markers by Western Bloting.The mechanism of HIF-2α promoting cancer cell migration and invasion ability was analyzed by Western bloting detecting EMTrelated indices.Results：When the plasmid of over-expression was transfected into hepatoma cells,the expression levels of HIF-2α mRNA and protein levels both increased（P〈0.05）.The Transwell movement experiments showed that after the high expression of HIF-2α,the number of transmembrane cells increased significantly（P〈0.01）.The scratch test results also showed that the high expression of HIF-2α enhanced the movement ability of cells.The in vivo invasion experiment showed that the high expression of HIF-2α increased the lung metastasis tumor size in mice significantly.Western bloting results showed that HIF-2α can promote the protein expression of mesenchymal cells-associated markers.Conclusions：HIF-2α might improve the movement and invasion ability of hepatocellular carcinoma cells（HCC）by promoting their epithelial-mesenchymal transition（EMT）.

关 键 词：HIF-2α 肝癌细胞 运动 侵袭 

分 类 号：R735.7[医药卫生—肿瘤]