Publication date: 5 February 2018
Source:Gene, Volume 642
Author(s): Chunchun Zhao, Zhen Xu, Zijie Wang, Chuanjian Suo, Jun Tao, Zhijian Han, Min Gu, Ruoyun Tan
BackgroundChronic allograft dysfunction (CAD) is characterized by allograft kidney interstitial fibrosis, the underlying mechanism of which is unclear. Our aim was to elucidate the role and mechanism of TNF-α-induced epithelial-to-mesenchymal transition (EMT) in transplant kidney tubular interstitial fibrosis.MethodsHuman kidney tissues from normal volunteers and CAD patients were assessed using periodic acid-Schiff, Masson trichrome and immunohistochemical staining. mRNA and protein expression of E-cadherin, α-smooth muscle actin (SMA) and fibronectin(FN) in renal proximal tubule epithelial (HK-2) cells after treatment with TNF-α under different conditions were assessed using western blot and qRT-PCR analysis. Cell motility and migration were assessed using wound healing and transwell assays. Expression of Smurf2 and TNF-α-signaling pathway-related proteins in HK-2 cells treated with TNF-α was detected by western blotting. E-cadherin and α-SMA expression was also assessed in Smurf2 plasmid-transfected or Smurf2 siRNA-treated HK-2 cells.ResultsThe expression of TNF-α, Smurf2, α-SMA, and fibronectin was significantly upregulated, while the expression of E-cad was downregulated in the CAD group compared with the normal group. The in vitro results showed that TNF-α remarkably upregulated the expression of Smurf2, α-SMA and fibronectin and downregulated the expression of E-cadherin in HK-2 cells and enhanced motility and migration in HK-2 cells. Overexpression of Smurf2 could promote the expression of α-SMA and inhibit the expression of E-cad, whereas knockdown of Smurf2 expression reversed TNF-α-induced upregulation of α-SMA and prohibited the reduction of E-cad expression. Furthermore, TNF-α-induced Smurf2 expression promoted EMT through the Akt signaling pathway.ConclusionsTNF-α induced EMT via the TNF-α/Akt/Smurf2 signaling pathways, and it may play a role in aggravating allograft kidney interstitial fibrosis in CAD patients.
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