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CJAP ›› 2017, Vol. 33 ›› Issue (4): 346-350.doi: 10.12047/j.cjap.5477.2017.084

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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)

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

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