Roles of layilin in TNF-α-induced epithelial-mesenchymal transformation of renal tubular epithelial cells

Biochem Biophys Res Commun. 2015 Nov 6;467(1):63-9. doi: 10.1016/j.bbrc.2015.09.121. Epub 2015 Sep 26.

Abstract

Background: Tumor necrosis factor (TNF)-α is suggested to induce epithelial-mesenchymal transformation (EMT) of renal tubular epithelial cells that possibly exacerbates renal interstitial fibrosis in glomerulonephritis (GN). We here investigated whether layilin (LAYN), a c-type lectin-homologous protein, was involved in the EMT process.

Methods: Expression of LAYN was investigated in kidneys of mice administered with TNF-α and in a clear cell renal carcinoma cell line of KMRC-1 stimulated with TNF-α by quantitative polymerase chain reaction (qPCR) and/or western blotting. Expression of LAYN was assessed immunohistochemically in renal biopsy samples of patients with various types of GN. Changes of EMT markers and cell morphology by TNF-α and transforming growth factor (TGF)-β in LAYN-knocked down KMRC-1 cells were investigated by qPCR and immunocytochemistry.

Results: Administration of TNF-α increased expression of LAYN in renal tubular epithelia in mice. TNF-α but not TGF-β increased expression of LAYN in KMRC-1 cells. Renal biopsy samples from the patients with GN showed high expression of LAYN in tubular epithelial cells. TNF-α induced up-regulation of vimentin, down-regulation of E-cadherin, and fibroblast-like morphological change in KMRC-1 cells, indicating occurrence of EMT. These changes were not observed in the LAYN-knocked down cells. In contrast, similarly occurred TGF-β-induced EMT was not affected by the LAYN knockdown.

Conclusion: Our data indicate that LAYN is involved in the TNF-α-induced EMT of renal tubular epithelial cells. LAYN may play roles in the generation of renal interstitial fibrosis in GN via TNF-α-induced EMT.

Keywords: Epithelial-mesenchymal transformation; Glomerulonephritis; Layilin; Renal tubular epithelium; TNF-α.

MeSH terms

  • Adult
  • Animals
  • Carrier Proteins / metabolism*
  • Cell Line
  • Epithelial Cells / cytology
  • Epithelial Cells / drug effects
  • Epithelial Cells / metabolism
  • Epithelial-Mesenchymal Transition / drug effects
  • Epithelial-Mesenchymal Transition / physiology*
  • Female
  • Gene Knockdown Techniques
  • Glomerulonephritis / metabolism
  • Humans
  • Kidney Tubules / cytology
  • Kidney Tubules / drug effects
  • Kidney Tubules / metabolism*
  • Lectins, C-Type / antagonists & inhibitors
  • Lectins, C-Type / genetics
  • Lectins, C-Type / metabolism*
  • Male
  • Membrane Glycoproteins / metabolism*
  • Mice
  • Mice, Inbred ICR
  • Tumor Necrosis Factor-alpha / metabolism*
  • Tumor Necrosis Factor-alpha / pharmacology
  • Young Adult

Substances

  • Carrier Proteins
  • LAYN protein, human
  • Lectins, C-Type
  • Membrane Glycoproteins
  • Tumor Necrosis Factor-alpha
  • layilin protein, mouse