[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.
点击复制

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()
分享到：

《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.

相似文献/References:

[1]陈耿臻,韩慧,许铭炎,等. 重组腺病毒ADV-TK 基因的构建及其对肝癌细胞的杀伤作用[J].南方医科大学学报,2010,(08):1887.
　CHEN Geng-zhen,HU Hui,XU Ming-yan,et al. Construction of recombinant adenovirus containing TK gene and its effect against human liver cancer cells[J].Journal of Southern Medical University,2010,(12):1887.
[2]凌志海,孙权权,张耀伟,等.吉西他滨增强肝癌HepG2细胞体外辐射敏感性的机制研究[J].南方医科大学学报,2011,(12):1993.
[3]陈建发,李宇华,陈引香,等.Apollon siRNA提高肝癌细胞化疗敏感性的实验研究[J].南方医科大学学报,2011,(10):1701.
[4]杨义明,杜钢军,林海红.沙利度胺治疗肝癌的实验研究[J].南方医科大学学报,2005,(08):925.
　YANG Yi-ming,DU Gang-jun,LIN Hai-hong.Experimental study of thalidomide for treatment of murine hepatocellular carcinoma[J].Journal of Southern Medical University,2005,(12):925.
[5]季锡清,李朝龙,杨进城,等.血流阻断的缺血预处理技术在肝癌切除术中的应用[J].南方医科大学学报,2004,(01):66.
　JI Xi-qing,LI Chao-long,YANG Jin-cheng,et al.Application of ischemic preconditioning before hepatic vascular exclusion for resection of hepatocellular carcinoma[J].Journal of Southern Medical University,2004,(12):66.
[6]王素珍,孟维静,安洪庆,等.基于倾向指数匹配法的肝癌病人疗效评价[J].南方医科大学学报,2012,(09):1234.
[7]肖芦山,邹雪晶,胡威,等.microRNA-107在肝癌中的表达及临床意义[J].南方医科大学学报,2016,(07):974.
[8]玉艳红,原彤彤,黄力毅,等.外周血单核细胞hFgl2蛋白表达与不同临床类型肝病的关系[J].南方医科大学学报,2013,(03):436.
[9]戴小珍,熊新,王兰,等.CXCR7-shRNA慢病毒载体对人肝癌细胞生长及侵袭能力的抑制作用[J].南方医科大学学报,2013,(07):994.
[10]曲凯,林婷,魏吉超,等.Cisplatin induces cell cycle arrest and senescence via upregulating P53 and P21 expression in HepG2 cells[J].南方医科大学学报,2013,(09):1253.

更新日期/Last Update: 1900-01-01