title: N-cadherin is overexpressed in Crohn's stricture fibroblasts and promotes intestinal fibroblast migration.
abstract: BACKGROUND
  Intestinal fibroblasts mediate stricture formation in Crohn's disease (CD). Transforming growth factor-β₁ (TGF-β₁) is important in fibroblast activation, while cell attachment and migration is regulated by the adhesion molecule N-cadherin. The aim of this study was to investigate the expression and function of N-cadherin in intestinal fibroblasts in patients with fibrostenosing CD.
  
  
  METHODS
  Intestinal fibroblasts were cultured from seromuscular biopsies from patients undergoing resection for terminal ileal fibrostenosing CD (n = 14) or controls patients (n = 8). N-cadherin expression was assessed using Western blot and quantitative reverse-transcription polymerase chain reaction (qRT-PCR). Fibroblasts were stimulated with TGF-β₁ and selective pathway inhibitors Y27632, PD98050, and LY294002 were used to examine the Rho/ROCK, ERK-1/2, and Akt signaling pathways, respectively. Cell migration was assessed using a scratch wound assay. N-cadherin was selectively overexpressed using a plasmid.
  
  
  RESULTS
  Fibroblasts from fibrostenosing CD express increased constitutive N-cadherin mRNA and protein and exhibit enhanced basal cell migration relative to those from directly adjacent normal bowel. Control fibroblasts treated with TGF-β₁ induced N-cadherin in a dose-dependent manner which was inhibited by Rho/ROCK and Akt pathway modulation. Control fibroblasts exhibited enhanced cell migration in response to treatment with TGF-β₁ or transfection with an N-cadherin plasmid.
  
  
  CONCLUSIONS
  Fibroblasts from strictures in CD express increased constitutive N-cadherin and exhibit enhanced basal cell migration. TGF-β₁ is a potent inducer of N-cadherin in intestinal fibroblasts resulting in enhanced cell migration. The TGF-β₁-mediated induction of N-cadherin may potentiate Crohn's stricture formation.
authors.affiliation: Department of Surgery, St. Vincent's University Hospital, Dublin, Ireland.
authors.forename: John P
authors.initials: JP
authors.lastname: Burke
authors.forename: Michael F
authors.initials: MF
authors.lastname: Cunningham
authors.forename: Catherine
authors.initials: C
authors.lastname: Sweeney
authors.forename: Neil G
authors.initials: NG
authors.lastname: Docherty
authors.forename: P Ronan
authors.initials: PR
authors.lastname: O'Connell
mesh_terms: D000328:Adult; D000704:Analysis of Variance; D015820:Cadherins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D002465:Cell Movement / Q000502:physiology; D002478:Cells, Cultured; D003251:Constriction, Pathologic / Q000378:metabolism / Q000473:pathology; D003424:Crohn Disease / Q000378:metabolism* / Q000473:pathology*; D005260:Female; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D005355:Fibrosis; D006801:Humans; D007082:Ileum / Q000473:pathology*; D008297:Male; D012333:RNA, Messenger / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology; D015854:Up-Regulation / Q000235:genetics
chemical_list: D015820:Cadherins; D012333:RNA, Messenger; D053773:Transforming Growth Factor beta1