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NDRG2 knockdown promotes fibrosis in renal tubular epithelial cells through TGF-β1/Smad3 pathway

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Abstract

Renal fibrosis is a common pathological pathway of various chronic kidney diseases progressing to end-stage renal disease and is characterized by tubular atrophy, fibroblast/myofibroblast activation and excessive deposition of extracellular matrix (ECM). N-Myc downstream-regulated gene-2 (NDRG2) is reported to be associated with liver fibrosis in rats. However, the biological function of NDRG2 in renal fibrosis remains unclear. Therefore, we investigate the effect of NDRG2 on renal fibrosis and the underlying mechanism of NDRG2 in TGF-β1-induced renal tubular epithelial cells (HK-2). Our results show that TGF-β1 down-regulates NDRG2 mRNA and protein expression in HK-2 cells. Moreover, NDRG2 knockdown dramatically reduces the TGF-β1-induced protein and mRNA of E-cadherin and increases the TGF-β1-induced protein and mRNA expression level of α-SMA, Vimentin, Snail, Col-I, Col-III and FN; this is reversed by NDRG2 overexpression. Furthermore, NDRG2 silencing significantly increases the phosphorylation level of Smad3 (p-Smad3), which is decreased by NDRG2 overexpression, although these have no effect on the protein expression of p-Smad2 and Smad7. In addition, SIS3, a specific inhibitor of Smad3 phosphorylation, partly reverses the effect of NDRG2 knockdown on the protein and mRNA expression of epithelial-mesenchymal transition (EMT) markers and ECM components in TGF-β1-induced HK-2 cells. Taken together, our results indicate that NDRG2 knockdown promotes renal fibrosis through its effect on the protein and mRNA expression of EMT markers and ECM components by regulating the downstream Smad3 signaling pathway in renal tubular epithelial cells. Modulation of NDRG2 expression might provide a new therapy for renal fibrosis.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (nos. 81570685, 81100464, and 81200883), the Youth Foundation of the First Affiliated Hospital of Zhengzhou University for Doctor of Medicine, a General Financial Grant from the China Postdoctoral Science Foundation (no. 2012 M521410), the General Scientific and Technological project of Henan Province (201702031, 201702015), the Foundation of Henan Educational Committee (12A320061, 14A320097), and the Scientific and Technological Innovation Project of Zhengzhou City (131PCXTD627).

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Authors and Affiliations

  1. Department of Urology, Zhengzhou Key Laboratory for Molecular Biology of Urological Tumor Research, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, 450052, People’s Republic of China

    Zhibo Jin, Chaohui Gu, Zhankui Jia & Jinjian Yang

  2. Zhengzhou Key Laboratory for Molecular Biology of Urological Tumor Research, Henan Institute of Urology, Zhengzhou, Henan, 450052, People’s Republic of China

    Zhibo Jin, Chaohui Gu, Zhankui Jia & Jinjian Yang

  3. Department of Pediatrics, Zhengzhou Key Laboratory for Molecular Biology of Urological Tumor Research, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, 450052, People’s Republic of China

    Fengyan Tian

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  1. Zhibo Jin
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  3. Fengyan Tian
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  4. Zhankui Jia
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Contributions

Zhibo Jin and Chaohui Gu performed the experiments, analyzed the data and drafted the manuscript. Fengyan Tian assisted with cell culture, plasmid construction and transfections. Zhankui Jia assisted with gene and protein detection. Jinjian Yang conceived and designed the study and revised the manuscript.

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Correspondence to Jinjian Yang.

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All authors declare no competing interests.

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Zhibo Jin and Chaohui Gu contributed equally to this work.

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Jin, Z., Gu, C., Tian, F. et al. NDRG2 knockdown promotes fibrosis in renal tubular epithelial cells through TGF-β1/Smad3 pathway. Cell Tissue Res 369, 603–610 (2017). https://doi.org/10.1007/s00441-017-2643-7

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  • DOI: https://doi.org/10.1007/s00441-017-2643-7

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