N-cadherin is overexpressed in Crohn's stricture fibroblasts and promotes intestinal fibroblast migration

Inflamm Bowel Dis. 2011 Aug;17(8):1665-73. doi: 10.1002/ibd.21543. Epub 2011 Feb 1.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Analysis of Variance
  • Cadherins / genetics
  • Cadherins / metabolism*
  • Cadherins / physiology
  • Cell Movement / physiology
  • Cells, Cultured
  • Constriction, Pathologic / metabolism
  • Constriction, Pathologic / pathology
  • Crohn Disease / metabolism*
  • Crohn Disease / pathology*
  • Female
  • Fibroblasts / drug effects
  • Fibroblasts / metabolism*
  • Fibroblasts / physiology
  • Fibrosis
  • Humans
  • Ileum / pathology*
  • Male
  • RNA, Messenger / metabolism*
  • Signal Transduction / physiology
  • Transforming Growth Factor beta1 / pharmacology
  • Up-Regulation / genetics

Substances

  • Cadherins
  • RNA, Messenger
  • Transforming Growth Factor beta1