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

• Published in: Inflammatory bowel diseases 2011-08, Vol.17 (8), p.1665-1673
• Main Authors: Burke, John P., Cunningham, Michael F., Sweeney, Catherine, Docherty, Neil G., O'Connell, P. Ronan
• Format: Article
• Language: English
• Subjects: Adult; AKT protein; Analysis of Variance; Basal cells; Biopsy; Cadherins - genetics; Cadherins - metabolism; Cadherins - physiology; Cell activation; Cell adhesion; Cell adhesion molecules; Cell migration; Cell Movement - physiology; Cells, Cultured; Constriction, Pathologic - metabolism; Constriction, Pathologic - pathology; Crohn Disease - metabolism; Crohn Disease - pathology; Crohn's disease; Female; Fibroblasts; Fibroblasts - drug effects; Fibroblasts - metabolism; Fibroblasts - physiology; Fibrosis; Humans; Ileum - pathology; Inflammatory bowel diseases; intestinal fibroblasts; Intestine; Male; mRNA; N-Cadherin; Plasmids; Polymerase chain reaction; RNA, Messenger - metabolism; Signal transduction; Signal Transduction - physiology; Stenosis; Transfection; Transforming Growth Factor beta1 - pharmacology; Up-Regulation - genetics; Western blotting; Wounds
• ISSN: 1078-0998; 1536-4844; 1536-4844
• DOI: 10.1002/ibd.21543

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. (Inflamm Bowel Dis 2011;)