南方医科大学学报Issue(12)：1743-1747,5.DOI:10.3969/j.issn.1673-4254.2014.12.07
慢病毒介导shRNA靶向下调Cx26表达对人高侵袭性肝癌细胞上皮间质转化及侵袭的影响
Lentivirus-mediated shRNA interference of Cx26 suppresses epithelial mesenchymal transi-tion and invasion of highly invasive hepatocellular carcinoma cells in vitro
摘要
Abstract
Objective To explore the effect of lentivirus-mediated shRNA interference of Cx26 on epithelial-mesenchymal transition (EMT) and invasion of highly invasive human hepatocellular carcinoma cells in vitro. Methods SK-Hep-1 cells were infected with the lentivirus for delivering Cx26 shRNA, and the stably transfected cells were selected by puromycin. The interference efficiency of shRNA-Cx26 was assessed with real-time PCR and Western blotting. The morphological changes of the transfected SK-Hep-1 cells were observed microscopically, and the protein expressions of E-cadherin and vimentin were detected using Western blotting. The effect of Cx26 interference on the invasiveness of SK-Hep-1 cells was determined by Transwell invasion assay. Results Compared with SK-Hep-1 cells infected with empty EGFP vector and uninfected cells, the cells transfected with shRNA-Cx26 showed significantly reduced mRNA and protein expressions of Cx26 (P<0.01), which resulted in obvious morphological conversion from mesenchymal cells to epithelial cells. shRNA-Cx26-transfected cells showed significantly increased E-cadherin protein expression (P<0.01) but decreased vimentin expression (P<0.01) with obviously attenuated invasive ability in vitro (P<0.01). Conclusion Targeted down-regulation of Cx26 expression can inhibit the EMT and invasion of SK-Hep-1 cells in vitro.关键词
缝隙连接蛋白26/肝癌/上皮间质转化/侵袭/shRNAKey words
connexin26/hepatocellular carcinoma/epithelial-mesenchymal transition/invasion/short hairpin RNA引用本文复制引用
阳洁,覃贵慧,陈军泽..慢病毒介导shRNA靶向下调Cx26表达对人高侵袭性肝癌细胞上皮间质转化及侵袭的影响[J].南方医科大学学报,2014,(12):1743-1747,5.基金项目
国家自然科学基金（81260324）；教育部高等学校博士学科点专项科研基金新教师类（20124503120008）；广西教育厅高校科研项目（201106LX117）；广西医科大学青年科学基金（GXMUYSF14） Supported by National Natural Science Foundation of China (81260324) （81260324）
