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  • Title: VEGF suppresses epithelial-mesenchymal transition by inhibiting the expression of Smad3 and miR‑192, a Smad3-dependent microRNA.
    Author: Hong JP, Li XM, Li MX, Zheng FL.
    Journal: Int J Mol Med; 2013 Jun; 31(6):1436-42. PubMed ID: 23588932.
    Abstract:
    Transforming growth factor-β1 (TGF-β1)‑induced epithelial‑mesenchymal transition (EMT) is one of the important cellular and molecular mechanisms involved in renal fibrosis. Smad3 and miR-192 (a Smad3-dependent microRNA) are involved in TGF-β1-mediated EMT. Vascular endothelial growth factor (VEGF) is a renal tubular epithelial survival factor. Therefore, in the present study, we investigated the role of Smad3 and miR‑192 in the effects of VEGF on TGF‑β1‑mediated tubular EMT. A human kidney cortex (HKC) cell line stably overexpressing VEGF (HKC-SOEV) was established. The normal HKC cells and HKC‑SOEV cells were treated with TGF-β1 (5 µg/l) or/and LY294002 (20 µmol/l) for 24 and 48 h (LY294002 blocks the effect of VEGF). The protein expression of Smad2, Smad3, Smad4 and phosphorylated Smad3 (p‑Smad3) were measured by western blot analysis. The expression of Smad3 and miR-192 was determined by real‑time PCR. E-cadherin and α-smooth muscle actin (α-SMA) expression was detected by western blot analysis and laser scanning confocal microscopy (LSCM). TGF-β1 was found to induce the expression of α-SMA in the HKC cells. TGF-β1 also induced Smad3, miR-192 and p-Smad3 expression, but suppressed E‑cadherin expression. However, in the HKC-SOEV cells, the expression levels of α-SMA, Smad3, miR-192 and p‑Smad3 upon TGF-β1 stimulation were significantly reduced. In these cells, the suppressive effect of TGF-β1 on E‑cadherin was also reduced. Importantly, treatment with LY294002 significantly diminished the effect of VEGF. VEGF suppressed Smad3 and miR‑192, and subsequently inhibited EMT induced by TGF-β1.
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