N-cadherin is overexpressed in Crohn's stricture fibroblasts and promotes intestinal fibroblast migration
✍ Scribed by John P. Burke; Michael F. Cunningham; Catherine Sweeney; Neil G. Docherty; P. Ronan O'Connell
- Publisher
- John Wiley and Sons
- Year
- 2011
- Tongue
- English
- Weight
- 636 KB
- Volume
- 17
- Category
- Article
- ISSN
- 1078-0998
No coin nor oath required. For personal study only.
✦ Synopsis
Background: Intestinal fibroblasts mediate stricture formation in Crohn's disease (CD). Transforming growth factor-b 1 (TGF-b 1 ) 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 ¼ 14) or controls patients (n ¼ 8). N-cadherin expression was assessed using Western blot and quantitative reverse-transcription polymerase chain reaction (qRT-PCR). Fibroblasts were stimulated with TGF-b 1 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-b 1 induced Ncadherin 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-b 1 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-b 1 is a potent inducer of N-cadherin in intestinal fibroblasts resulting in enhanced cell migration. The TGF-b 1mediated induction of N-cadherin may potentiate Crohn's stricture formation.