[1]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.
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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()
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《南方医科大学学报》[ISSN:1673-4254/CN:44-1627/R]

卷:
期数:
2014年12期
页码:
1743
栏目:
出版日期:
2014-12-20

文章信息/Info

Title:
Lentivirus-mediated shRNA interference of Cx26 suppresses epithelial mesenchymal
transition and invasion of highly invasive hepatocellular carcinoma cells in vitro
作者:
阳洁覃贵慧陈军泽
关键词:
缝隙连接蛋白26肝癌上皮间质转化侵袭shRNA
Keywords:
connexin26 hepatocellular carcinoma epithelial-mesenchymal transition invasion short hairpin RNA
摘要:
目的探讨慢病毒介导shRNA靶向下调缝隙连接蛋白26(Cx26)表达对人高侵袭性肝癌细胞上皮间质转化(EMT)及侵袭
的影响。方法用慢病毒介导Cx26-shRNA感染SK-Hep-1细胞并用嘌呤霉素筛选稳转细胞。荧光定量PCR和Western blot检
测干扰效率。光镜观察细胞形态改变。Western blot检测EMT上皮标志物E-cadherin、间质标志物vimentin的蛋白表达水平。
Transwell 侵袭实验检测细胞侵袭能力变化。结果成功建立稳定下调Cx26 表达的SK-Hep-1 细胞(shRNA-Cx26 组),与感染
EGFP空载体(shRNA-control组)及未感染的SK-Hep-1细胞(blank-control组)比较,其Cx26 mRNA和蛋白表达均明显降低(P<
0.01),间质细胞形态转变为上皮细胞形态,E-cadherin蛋白表达明显增多、vimentin蛋白表达明显减少(P<0.01),体外侵袭能力
也显著降低(P<0.01)。结论靶向下调Cx26表达能抑制人高侵袭性肝癌SK-Hep-1细胞的EMT和体外侵袭。
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.

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更新日期/Last Update: 1900-01-01