[1]周宝尚,张璟,陶光利.shRNA干扰β-catenin对TGF-β1诱导的肾小管上皮细胞转分化的作用[J].陆军军医大学学报(原第三军医大学学报),2011,33(20):2166-2171.
 Zhou Baoshang,Zhang Jing,Tao Guangli.Short hairpin RNA of β-catenin attenuates TGF-β1-induced epithelial mesenchymal transition in human proximal tubular epithelial cells[J].J Amry Med Univ (J Third Mil Med Univ),2011,33(20):2166-2171.
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shRNA干扰β-catenin对TGF-β1诱导的肾小管上皮细胞转分化的作用(/HTML )
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
33卷
期数:
2011年第20期
页码:
2166-2171
栏目:
论著
出版日期:
2011-10-30

文章信息/Info

Title:
Short hairpin RNA of β-catenin attenuates TGF-β1-induced epithelial mesenchymal transition in human proximal tubular epithelial cells
作者:
周宝尚张璟陶光利
第三军医大学新桥医院肾内科;重庆市第七人民医院肾内科
Author(s):
Zhou Baoshang Zhang Jing Tao Guangli
Department of Nephrology, Xinqiao Hospital, Third Military Medical University, Chongqing, 400037; Department of Nephrology, the 7th People’s Hospital of Chongqing, Chongqing, 400054, China
关键词:
β-连环蛋白短发卡RNA转化生长因子β1转分化肾小管纤维化
Keywords:
β-catenin short hairpin RNA transforming growth factor beta 1 epithelial-mesenchymal transition renal tubulointerstitialfibrosis
分类号:
R322.61;R394-33;R394.2
文献标志码:
A
摘要:
目的      构建特异性针对β-catenin基因的shRNA干扰载体,转染TGF-β1诱导的人近端肾小管上皮细胞株HKCs,研究其对肾小管上皮细胞转分化的影响。      方法      将构建的shRNA-β-catenin干扰载体转染TGF-β1诱导的HKC细胞,RT-PCR、Western blot检测β-catenin的mRNA及蛋白表达水平,筛选抑制效果最佳载体;RT-PCR、细胞免疫荧光、Western blot检测肾小管上皮细胞转分化相关蛋白E-cadherin、α-SMA的mRNA及蛋白表达水平。      结果      经酶切和测序鉴定,成功构建的shRNA-β-catenin表达载体转染HKC细胞,β-catenin在mRNA及蛋白表达水平均低于转染空白载体组(P<0.05),并筛选shRNA-β-catenin-1进入实验组;转染shRNA-β-catenin-1组细胞β-catenin在mRNA及蛋白表达水平均低于TGF-β1诱导组和空载体组(P<0.05),且β-catenin转核显著减弱(P<0.05);E-cadherin的mRNA及蛋白表达水平较TGF-β1诱导组和空载体组显著上调(P<0.05)、α-SMA的mRNA及蛋白表达水平较TGF-β1诱导组和空载体组显著下调(P<0.05)。      结论      运用shRNA-β-catenin干扰载体转染TGF-β1诱导的HKC细胞,能够抑制肾小管上皮细胞-间充质转分化。
Abstract:
Objective      To determine the effect of  β-catenin gene shRNA on the epithelial mesenchymal transition in the human proximal tubular epithelial cells (HKCs) induced by TGF-β1.       Methods      shRNA-β-catenin vector was constructed and then transfect into HKC cells induced by TGF-β1. RT-PCR and Western blot analysis were used to detect mRNA and protein expression levels of β-catenin to screen the best inhibitory vector. RT-PCR, immunofluorescence, and Western blot analysis were employed to detect mRNA and protein expression levels of renal tubular epithelial cell differentiation-related proteins, α-SMA and E-cadherin.       Results      The recombinant plasmid shRNA-β-catenin was confirmed by digestion and sequencing. After HKC cells were transfected with the plasmid, β-catenin expression at the mRNA and protein levels were significantly lower than the empty vector group (P<0.05). Plasmid shRNA-β-catenin-1 was screened, and transfected in HKC cells.β-catenin expression at the mRNA and protein levels were significantly lower in the cells transfected with shRNA-β-catenin-1 than in the cells with TGF-β1 inducement and the cells transfected with empty vector (P<0.05). In the shRNA-β-catenin-1 group, the amount of nuclear transferred β-catenin was significantly reduced compared with TGF-β1-induced group and the empty vector group (P<0.05), while E-cadherin mRNA and protein expression levels were significantly increased than the other 2 groups (P<0.05). On the contrast, compared with TGF-β1-induced group and the empty vector group, α-SMA mRNA and protein expression levels were significantly lower in the group (P<0.05).       Conclusion      shRNA-β-catenin vector attenuates epithelial mesenchymal transdifferentiation in human proximal tubular epithelial cells induced by TGF-β1.

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更新日期/Last Update: 2011-10-17