乙肝病毒X蛋白活化c-Src激酶诱导肝癌细胞发生上皮-间充质转化的机制  

Hepatitis B virus X protein induces epithelial-mesenchymal transition through c-Src activation in liver cancer cells

作　　者：杨世忠[1] 赵志远[2] 袁伟升[1] 陈耿[4] 张玉君[4] 张爱群[3] 董家鸿[3] 

机构地区：[1]解放军第401医院肝胆外科,青岛266071 [2]济南军区总医院第二门诊部,济南 [3]解放军总医院肝胆外科,北京 [4]第三军医大学西南医院肝胆外科,重庆

出　　处：《中华肝胆外科杂志》2015年第3期200-203,共4页Chinese Journal of Hepatobiliary Surgery

基　　金：国家传染病科技重大专项（2012ZXl0002-017）;中国博士后科学基金资助（200902681）

摘　　要：目的 探讨在乙肝病毒X蛋白（HBx）诱导肝癌细胞上皮-间充质转化（EMT）过程中癌基因c-Src的作用.方法 通过HBx基因转染诱导SMMC-7721肝癌细胞发生上皮-间充质转化,量化分析实验组和对照组细胞c-Src激酶的活化情况.抑制c-Src活性,通过实时PCR、蛋白印迹、免疫细胞化学等方法评价c-Src活化对上皮-间充质转化的影响.结果 发生上皮-间充质转化的SMMC-7721细胞中c-Src活化显著增加.给予c-Src激酶抑制剂PP2后,细胞由梭形恢复原来的上皮表型,而给予安慰剂PP3细胞形态无明显变化.蛋白印迹、免疫细胞化学检测进一步证实,抑制c-Src活化能逆转HBx基因转染诱导的上皮-间充质转化,使细胞再次发生间叶-上皮样转化,即上皮标志物（E-cadherin、α-catenin）表达显著上调,间叶标志物（N-cadherin,Vimentin,Fibronectin）表达显著减少.结论 HBx蛋白诱导SMMC-7721肝癌细胞发生上皮-间充质转化时,c-Src活化发挥了关键作用.Objective To examine the role of c-Src activation in hepatitis B virus X （HBx） protein induced epithelial-mesenchymal transition （EMT） in liver cancer.Methods SMMC-7721 liver cancer cells were transfected with HBx gene to induce EMT and the activated c-Src expression was evaluated by Western blot.Both the morphological changes and the epithelial and mesenchymal markers expression （real-time PCR,western blot and immunocytochemistry） of HBx-transfected SMMC-7721 cell treated by c-Src kinase inhibitor PP2 and negative control PP3 were observed and compared,respectively.Results The activated c-Src expression in HBx gene transfected SMMC-7721 cells was significantly increased compared to that in mock transfected cells,c-Src kinase inhibitor PP2 could enable the HBx-transfected SMMC-7721 cells to transmit from spindle-like shape to original epithelial morphology.Western blot and immunocytochemistry confirmed that the expression of epithelial markers and mesenchymal markers almost returned to the levels of parental cells,indicating the mesenchymal-epithelial transition.Conclusions c-Src activation plays a key role in the process of EMT induced by HBx protein in SMMC-7721 cells.

关 键 词：乙肝病毒X蛋白 上皮-间充质转化 肝癌细胞 c-Src激酶 

分 类 号：R-322[医药卫生]