E-Cadherin Expression Is Regulated by miR-192/215 by a Mechanism That Is Independent of the Profibrotic Effects of Transforming Growth Factor-β

Abstract

OBJECTIVEIncreased deposition of extracellular matrix (ECM) within the kidney is driven by profibrotic mediators including transforming growth factor-β (TGF-β) and connective tissue growth factor (CTGF). We investigated whether some of their effects may be mediated through changes in expression of certain microRNAs (miRNAs).RESEARCH DESIGN AND METHODSProximal tubular cells, primary rat mesangial cells, and human podocytes were analyzed for changes in the expression of key genes, ECM proteins, and miRNA after exposure to TGF-β (1–10 ng/μl). Tubular cells were also infected with CTGF-adenovirus. Kidneys from diabetic apoE mice were also analyzed for changes in gene expression and miRNA levels.RESULTSTGF-β treatment was associated with morphologic and phenotypic changes typical of epithelial-mesenchymal transition (EMT) including increased fibrogenesis in all renal cell types and decreased E-cadherin expression in tubular cells. TGF-β treatment also modulated the expression of certain miRNAs, including decreased expression of miR-192/215 in tubular cells, mesangial cells, which are also decreased in diabetic kidney. Ectopic expression of miR-192/215 increased E-cadherin levels via repressed translation of ZEB2 mRNA, in the presence and absence of TGF-β, as demonstrated by a ZEB2 3′-untranslated region luciferase reporter assay. However, ectopic expression of miR-192/215 did not affect the expression of matrix proteins or their induction by TGF-β. In contrast, CTGF increased miR-192/215 levels, causing a decrease in ZEB2, and consequently increased E-cadherin mRNA.CONCLUSIONSThese data demonstrate the linking role of miRNA-192/215 and ZEB2 in TGF-β/CTGF–mediated changes in E-cadherin expression. These changes appear to occur independently of augmentation of matrix protein synthesis, suggesting that a multistep EMT program is not necessary for fibrogenesis to occur.

The transcriptome of nitrofen-induced pulmonary hypoplasia in the rat model of congenital diaphragmatic hernia

“…From our study, we identified a nitrofen-induced upregulation of rno-miR-296-3p. MiR-192/215 has been shown to mediate changes in E-cadherin expression via targeting the ZEB2 family of transcriptional repressors downstream of TGF-β (29). In addition, our study showed downregulation of members of the miR-200 family (rno-miR-200c-3p, rno-miR-200c-5p, and rno-miR-200b-5p) upon nitrofen treatment by >twofold relative to the control.…”

Characterization of gene expression and activated signaling pathways in solid-pseudopapillary neoplasm of pancreas

“…A recent study demonstrated that the miRNA-200 family (miR-141, miR-200a, b, c, and miR-429) and miR-192/215 inhibit E-cadherin expression by targeting ZEB1 and ZEB2. [33][34][35][36][37] These previous reports suggest that solid-pseudopapillary neoplasm-specific downregulation of the miR-200 family and miR-192/215 targets components of the Wnt/b-catenin, Hedgehog, androgen receptor signaling pathways as well as genes associated with epithelial-mesenchymal transition. These effects ultimately result in decreased expression of epithelial markers.…”

microRNAs in Diabetic Kidney Disease

“…Wang et al also reported that decreased expression of miR-192 and miR-215 by TGF-1 (10 ng/mL for 72 h) in primary rat MCs and TEC, and in the renal cortex of apolipoprotein E knockout mice at 10 weeks of diabetes [61]. These observed differences in murine models may be due to the different models and time points that were used [56,61]. Reduced miR-192 and miR-215, which target Zeb2, are involved in TGF-/CTGFmediated changes in E-cadherin expression, demonstrating that miR-192/215 may not affect fibrosis by directly altering the expression of fibrotic markers and ECM proteins [61].…”

“…In HK-2 cells, a human proximal TEC line, TGF-1 (10 ng/mL for 96 h) reduces the expression of miR-192 and zinc finger E-box binding homeobox 1 (Zeb1) accompanied by a reduction of E-cadherin and upregulation of zinc finger E-box binding homeobox 2 (Zeb2), PAI-I, and vimentin [60]. Wang et al also reported that decreased expression of miR-192 and miR-215 by TGF-1 (10 ng/mL for 72 h) in primary rat MCs and TEC, and in the renal cortex of apolipoprotein E knockout mice at 10 weeks of diabetes [61]. These observed differences in murine models may be due to the different models and time points that were used [56,61].…”

“…These observed differences in murine models may be due to the different models and time points that were used [56,61]. Reduced miR-192 and miR-215, which target Zeb2, are involved in TGF-/CTGFmediated changes in E-cadherin expression, demonstrating that miR-192/215 may not affect fibrosis by directly altering the expression of fibrotic markers and ECM proteins [61].…”