Lentivirus-mediated shRNA interference of Cx26 suppresses epithelial mesenchymal transi-tion and invasion of highly invasive hepatocellular carcinoma cells in vitro
10.3969/j.issn.1673-4254.2014.12.07
- VernacularTitle: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
- Author:
Jie YANG
1
;
Guihui QIN
;
Junze CHEN
Author Information
1. 广西医科大学 药学院药理学教研室
- Keywords:
connexin26;
hepatocellular carcinoma;
epithelial-mesenchymal transition;
invasion;
short hairpin RNA
- From:
Journal of Southern Medical University
2014;(12):1743-1747
- CountryChina
- Language:Chinese
-
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.
