title: Roles of layilin in TNF-α-induced epithelial-mesenchymal transformation of renal tubular epithelial cells.
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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Clinical Proteomics and Molecular Medicine, St. Marianna University Graduate School of Medicine, Kanagawa, Japan; Division of Nephrology and Hypertension, Department of Internal Medicine, St. Marianna University School of Medicine, Kanagawa, Japan.
authors.forename: Takayuki
authors.initials: T
authors.lastname: Adachi
authors.affiliation: Clinical Proteomics and Molecular Medicine, St. Marianna University Graduate School of Medicine, Kanagawa, Japan.
authors.forename: Mitsumi
authors.initials: M
authors.lastname: Arito
authors.affiliation: Clinical Proteomics and Molecular Medicine, St. Marianna University Graduate School of Medicine, Kanagawa, Japan.
authors.forename: Naoya
authors.initials: N
authors.lastname: Suematsu
authors.affiliation: Department of Anatomy, St. Marianna University School of Medicine, Kanagawa, Japan.
authors.forename: Atsuko
authors.initials: A
authors.lastname: Kamijo-Ikemori
authors.affiliation: Clinical Proteomics and Molecular Medicine, St. Marianna University Graduate School of Medicine, Kanagawa, Japan.
authors.forename: Kazuki
authors.initials: K
authors.lastname: Omoteyama
authors.affiliation: Clinical Proteomics and Molecular Medicine, St. Marianna University Graduate School of Medicine, Kanagawa, Japan.
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Sato
authors.affiliation: Disease Biomarker Analysis and Molecular Regulation, St. Marianna University Graduate School of Medicine, Kanagawa, Japan.
authors.forename: Manae S
authors.initials: MS
authors.lastname: Kurokawa
authors.affiliation: Clinical Proteomics and Molecular Medicine, St. Marianna University Graduate School of Medicine, Kanagawa, Japan.
authors.forename: Kazuki
authors.initials: K
authors.lastname: Okamoto
authors.affiliation: Japan Community Health Care Organization Tokyo Takanawa Hospital, Tokyo, Japan.
authors.forename: Kenjiro
authors.initials: K
authors.lastname: Kimura
authors.affiliation: Division of Nephrology and Hypertension, Department of Internal Medicine, St. Marianna University School of Medicine, Kanagawa, Japan.
authors.forename: Yugo
authors.initials: Y
authors.lastname: Shibagaki
authors.affiliation: Clinical Proteomics and Molecular Medicine, St. Marianna University Graduate School of Medicine, Kanagawa, Japan. Electronic address: t3kato@marianna-u.ac.jp.
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Kato
mesh_terms: D000328:Adult; D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism*; D002460:Cell Line; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects / Q000502:physiology*; D005260:Female; D055785:Gene Knockdown Techniques; D005921:Glomerulonephritis / Q000378:metabolism; D006801:Humans; D007684:Kidney Tubules / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D037181:Lectins, C-Type / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D008297:Male; D008562:Membrane Glycoproteins / Q000378:metabolism*; D051379:Mice; D008813:Mice, Inbred ICR; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism* / Q000494:pharmacology; D055815:Young Adult
chemical_list: D002352:Carrier Proteins; C546934:LAYN protein, human; D037181:Lectins, C-Type; D008562:Membrane Glycoproteins; D014409:Tumor Necrosis Factor-alpha; C115279:layilin protein, mouse
keywords: Epithelial-mesenchymal transformation; Glomerulonephritis; Layilin; Renal tubular epithelium; TNF-α