乙型肝炎病毒剪接特异性蛋白HBSP与TGFβ1诱导蛋白1相互作用促进TGFβ1诱导的肝癌细胞上皮间质转化
Hepatitis B spliced protein triggers TGFβ1-induced epithelial-mesenchymal transition via interaction with transforming growth factor beta-1-induced transcript 1 protein
摘要
Abstract
To investigate the TGFβ1-induced epithelial-mesenchymal transition (EMT) of Huh7 hepatoma cells caused by interaction of hepatitis B spliced protein (HBSP) with transforming growth factor beta-1-induced transcript 1 protein (TGFβ31I1),coding region of HBSP was cloned into lentiviral expression vector.Huh7 hepatoma cells were infected by recombinant lentivirus packaged in 293T cells.Stable cell lines expressing HBSP or control cells were selected by puromycin.Cells were incubated with 5 ng/mL TGFβ1 for 24 h,and observed under contrast-phase microspcope.Then the whole cell lysates were collected for western blot analysis using specific antibodies against EMT markers including E-cadherin,N-cadherin,Claudin-1 and β-catenin.To evaluate the effects of HBSP-TGFβ1I1 interaction on EMT,TGFβ1-induced EMT marker transition,as well as cell invasion and migration were explored after knocking down of TGFβ1I1 by siRNA.Results showed that Huh7 cell lines expressing HBSP (Huh7-HBSP flag-HIV) and control cell lines (Huh7-flag-HIV) were successfully established.Huh7-HBSP flag-HIV cells lost their pebble-like shape and tight cell-cell adhesion and transformed into the mesenchymal-like cells in the presence of TGFβ1.Decreased expression level of epithelial marker of E-cadherin,Claudin-1,β-catenin,increased expression level of mesenchymal marker of N-cadherin,and enhanced migration and invasion abilities were observed in Huh7-HBSP-flag-HIV cells as compared to the control cells.Moreover,the changes of EMT markers and metastasis abilities of Huh7-HBSP-flag-HIV cells could be reversed when TGFβ111 was knocked down by siRNA.In conclusion,HBSP could promote hepatoma cell migration and invasion by triggering EMT via interaction with TGFβ111.Our findings highlight new insights for HBSP-induced HCC progression.关键词
乙型肝炎病毒/RNA剪接/转化生长因子β1诱导蛋白1/上皮间质转化/侵袭Key words
hepatitis B virus/RNA splicing/transforming growth factor beta-1-induced transcript 1 protein/epithelial-mesenchymal transition/metastasis分类
医药卫生引用本文复制引用
陈婉南,黄俊高,梁菲菲,闫小利,轩丹丹,林旭..乙型肝炎病毒剪接特异性蛋白HBSP与TGFβ1诱导蛋白1相互作用促进TGFβ1诱导的肝癌细胞上皮间质转化[J].中国人兽共患病学报,2017,33(4):305-311,7.基金项目
国家自然科学基金青年基金项目(No.81201293) （No.81201293）
福建省卫生厅医学创新课题(No.2012-CX-14) （No.2012-CX-14）
福建省高校杰出青年科研人才培育计划(No.JA11104) （No.JA11104）
国家卫生和计划生育委员会共建科学研究基金-福建省卫生教育联合攻关计划(No.WKJ-FJ-29) （No.WKJ-FJ-29）
福建省高等学校新世纪优秀人才支持计划(No.JA13129) （No.JA13129）
福建省卫生系统中青年骨干人才培养项目(No.2014-ZQNZD-25)联合资助 Supported by the National Natural Science Foundation of China (No.81201293),the Fujian Provincial Medical Innovation Project (No.2012-CX-14),the Outstanding Young Researchers Training Plan of Fujian Province (No.JA11104),the Joint Research Program of Health and Planning Committee and Education Department of Fujian (No.WKJ-FJ-29),the New Century Talents Supporting Plan of Fujian Education Department (No.JA13129),and the Medical Elite Cultivation Program of Fujian (No.2014-ZQN-ZD-25) （No.2014-ZQNZD-25）
