首页  期刊介绍 征稿简则 编委会 期刊征订 广告服务 留言板 联系我们 English

中国应用生理学杂志 ›› 2017, Vol. 33 ›› Issue (4): 346-350.doi: 10.12047/j.cjap.5477.2017.084

• 研究论文 • 上一篇    下一篇

miR-21对TGF-β1诱导的肾小管上皮细胞间质转分化的影响

蔡月琴, 褚燕青, 朱科燕, 陈诚, 王德军   

  1. 浙江中医药大学 动物实验研究中心/比较医学研究所, 杭州 315700
  • 收稿日期:2016-07-21 修回日期:2017-01-30 出版日期:2017-07-28 发布日期:2018-06-19
  • 通讯作者: 王德军,Tel:0571-86613662;E-mail:wdj0369@126.com E-mail:wdj0369@126.com
  • 基金资助:
    浙江省自然科学基金资助项目(LQ14H290004);浙江中医药大学比较医学创新团队资助项目(XTD201301)

The influence of miR-21 on HK-2 EMT cells induced by TGF-beta 1

CAI Yue-qin, CHU Yan-qing, ZHU Ke-yan, CHEN Cheng, WANG De-jun   

  1. Zhejiang Chinese Medical University, Laboratory Animal Research Center/Comparative Medicine Institude, Hangzhou 310053, China
  • Received:2016-07-21 Revised:2017-01-30 Online:2017-07-28 Published:2018-06-19
  • Supported by:
    浙江省自然科学基金资助项目(LQ14H290004);浙江中医药大学比较医学创新团队资助项目(XTD201301)

摘要: 目的:观察miR-21在转化生长因子β1(TGF-β1)诱导的人肾小管上皮细胞(HK-2细胞)上皮间质转分化(EMT)中的作用,并探讨miR-21参与调控HK-2细胞EMT的可能靶点。方法:体外培养的HK-2细胞分为6组:正常对照组、转化生长因子β1(TGF-β1)模型组、miR-21 mimic阴性组、miR-21 mimic组、miR-21 inhibitor阴性组和miR-21 inhibitor组。细胞经4 ng/ml TGF-β1处理建立EMT模型,检测miR-21和EMT相关指标的表达变化,利用基因转染技术,将miR-21 mimic质粒或miR-21 inhibitor质粒转染经TGF-β1处理的HK-2细胞,使细胞过表达或抑制表达miR-21,在此基础上观察细胞EMT相关指标的变化以及磷酸酯酶(PTEN)基因的影响。结果:①与正常组相比,模型组的miR-21含量显著升高(P<0.05),上皮表型标志物E-cadherin的mRNA和蛋白表达水平均显著降低(P<0.01),间质表型标志物α平滑肌肌动蛋白(α-SMA) mRNA和蛋白水平也显著升高(P<0.05,P<0.01);②转染miR-21 mimic后,与miR-21 mimic阴性对照组相比,miR-21含量显著升高(P<0.01),PTEN、E-cadherin的mRNA和蛋白水平显著降低(P<0.05,P<0.01),α-SMA mRNA和蛋白水平显著升高(P<0.05,P<0.01);转染miR-21 inhibitor后,与miR-21 inhibitor阴性对照组相比,miR-21含量显著降低(P<0.01),PTEN、E-cadherin的mRNA和蛋白含量显著升高(P<0.05,P<0.01),α-SMA mRNA、蛋白水平显著降低(P<0.05,P<0.01)。结论:miR-21在TGF-β1诱导的HK-2细胞EMT发生中具有重要作用,并且可能通过靶基因PTEN参与EMT相关分子的表达调控。

关键词: 微小RNA-21, HK-2细胞, 转化生长因子-β1, 上皮间质转分化

Abstract: Objective: To investigate the effect of the miR-21 and its target mRNA in renal tubular epithelial mesenchymal transformation (EMT) model induced by transformation growth factor-β1(TGF-β1) in human renal tubular epithelial (HK-2) cells.Methods: HK-2 cells were divided into 6 groups:normal control group, TGF-β1 group, miR-21 mimic negative group, miR-21 mimic group, miR-21 inhibitor negative group and miR-21 inhibitor group. EMT model was established in HK-2 cells induced by 4 ng/ml TGF-β1. The level of miR-21, the mRNA and protein expression of EMT related factors were detected. MiR-21 mimic plasmid and miR-21 inhibitor plasmid were transfected into HK-2 cells that treated with TGF-β1 respectively using liposome transfection technique. Observe the impact of overexpression or inhibition expression of miR-21 on the mRNA and protein expression of EMT related factors and PTEN.Results: ①Compared with the normal group, the level of miR-21 was significantly increased in model group (P<0.05), the mRNA and protein expression levels of epithelial cells marker E-cadherin was significantly decreased (P<0.01), while the mRNA and protein levels of mesenchymal cells marker α-SMA was significantly increased (P<0.05,P<0.01). ②Compared with the miR-21 mimic negative group, the level of miR-21 in miR-21 mimic group increased significantly (P<0.01), the mRNA and protein expression levels of PTEN and E-cadherin decreased significantly (P<0.05,P<0.01), the mRNA and protein levels of α-SMA increased significantly (P<0.05,P<0.01). Compared with the miR-21 inhibitor negative control group, the level of miR-21 in miR-21 inhibitor group decreased significantly (P<0.01), the mRNA and protein expression levels of PTEN and E-cadherin increased significantly (P<0.05,P<0.01), the mRNA and protein levels of α-SMA decreased significantly (P<0.05,P<0.01).Conclusion: MiR-21 may play an important role in EMT induced by TGF-β1 in HK-2 cells and regulate the expression of EMT related factors its target gene PTEN.

Key words: microRNA-21, HK-2 cells, transformation growth factor-β1, epithelial mesenchymal transformation

版权所有 © 2015 《中国应用生理学杂志》编辑部
京ICP备16058274号-1
地址:天津市和平区大理道1号,邮编:300050  电话:022-23909086  E-mail:editor@cjap.ac.cn
本系统由北京玛格泰克科技发展有限公司设计开发 技术支持:support@magtech.com.cn