基础医学与临床 ›› 2010, Vol. 30 ›› Issue (10): 1029-1036.

• 研究论文 • 上一篇    下一篇

VEGF抑制HK2细胞上皮-间充质转化过程中NOTCH信号系统的变化

张颖娟 连耀国 李雪梅 苏颖 郑法雷   

  1. 中国医学科学院 北京协和医学院 北京协和医院 中国医学科学院 北京协和医学院 北京协和医院 肾内科 中国医学科学院 北京协和医学院 北京协和医院 肾内科 中国医学科学院 北京协和医学院 北京协和医院 肾内科
  • 收稿日期:2010-06-12 修回日期:2010-07-12 出版日期:2010-10-05 发布日期:2010-10-05
  • 通讯作者: 郑法雷

Inhibition of TGF-β1-Induced Epithelial-mesenchymal transition (EMT) by VEGF in HK2 cells is Related to Down-regulations of JAGGED-1and NOTCH1 In HK2 Cells

Ying-juan ZHANG, Yao-guo LIAN, Xue-mei LI, Ying SU, Fa-lei ZHENG   

  1. Peking Union Medical College Hospital, PUMC Dept. of Nephrology, Peking Union Medical College Hospital, PUMC & CAMS
  • Received:2010-06-12 Revised:2010-07-12 Online:2010-10-05 Published:2010-10-05
  • Contact: Fa-lei ZHENG

摘要: 目的 探讨应用血管内皮生长因子(VEGF)干预肾小管上皮-间充质转化(EMT)过程中NOTCH信号系统的变化及其意义。方法 1.体外培养的人肾小管上皮细胞(HK2)分为转化生长因子-β1(TGF-β1,5μg/L)单独作用组(T组)、TGF-β1(5μg/L)与VEGF(100μg/L)共同作用组(T+V组),RT-PCR 及Western blot方法检测不同作用时间点JAGGED-1、α-平滑肌肌动蛋白(α-SMA)的表达。2. TGF-β1与不同浓度VEGF165(0.1、1、10和100μg/L)共同作用细胞24h,激光共聚焦显微镜(CFMS)、RT- PCR及Western blot检测E-cadherin、α-SMA、JAGGED-1、NOTCH1表达。 结果 1.作用后24和48h,T+V组α-SMA和JAGGED-1蛋白表达均明显低于T组(P<0.05)。2.同TGF-β1单独作用组比较,VEGF165(100 μg/L)与TGF-β1共同作用组E-cadherin表达明显增强,α-SMA、 JAGGED-1及NOTCH1表达均明显降低(P<0.05)。结论 VEGF可抑制TGF-β1诱导HK2细胞发生EMT,并同时下调EMT过程中JAGGED-1、NOTCH1分子表达,提示该效应可能是VEGF抑制EMT的机制之一。

关键词: 血管内皮生长因子, 肾小管上皮-间充质细胞转化, Jagged-1, Notch1

Abstract: Objective To examine the effect of VEGF165 on expressions of JAGGED-1and NOTCH1 and the relationship between these expressions and epithelial- mesenchymal transition (EMT) induced by TGF-β1. Methods Cultured HK2 cells were incubated with TGF-β1(5μg/L) in the absence (group T) or presence (group T+V) of VEGF165 (100μg/L),respectively. Expressions of α-SMA and JAGGED-1mRNA were assessed with RT-PCR and the protein expressions were assessed with Western Blot at different time points,respectively. In the other experiment, HK2 cells were divided into four groups: negative control, incubated with TGF-β1(5μg/L) along, with VEGF165 (100μg/L) along, or with TGF-β1(5μg/L) and VEGF165 at different concentrations (0.1, 1, 10, 100μg/L) for 24 hours. Confocal microscope was used to detect expressions of α-SMA and E-cadherin. Protein and mRNA expressions of α-SMA, JAGGED-1and NOTCH1 were assessed with RT-PCR and Western Blot, respectively. Results Protein expression of both α-SMA and JAGGED-1decreased significantly in group T+V at 24h and 48h when compared to group T (P<0.05). VEGF165 reversed TGF β1-induced inhibition of E-cadherin and promotion of α-SMA, JAGGED-1and NOTCH1 significantly (P<0.05). Conclusions The results of the present study indicated that VEGF165 may partially inhibits TGFβ1-induced EMT of HK2 cells in vitro, which is probably contributed to the repression of JAGGED-1and NOTCH1 expressions. Further study is still needed.

Key words: Vascular endothelial growth factor, Epithelial-mesenchymal transition, TGF-β1, Notch1, Jagged-1