N-Cadherin Is Overexpressed in Crohn's Stricture Fibroblasts and Promotes Intestinal Fibroblast Migration
N-Cadherin Is Overexpressed in Crohn's Stricture Fibroblasts and Promotes Intestinal Fibroblast Migration
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DOI：
10.1002/ibd.21543
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发表时间：
2011-08-01
影响因子：
4.9
通讯作者：
O'Connell, P. Ronan
中科院分区：
文献类型：
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作者：
Burke, John P.;Cunningham, Michael F.;O'Connell, P. Ronan
背景：肠成纤维细胞介导了克罗恩病(CD)狭窄的形成。转化生长因子-β(1)在成纤维细胞的活化中起重要作用，而细胞的黏附和迁移受黏附分子N-钙粘附素的调节。本研究的目的是探讨N-钙粘附素在纤维狭窄CD患者肠成纤维细胞中的表达和功能。方法：取14例回肠末端狭窄患者和8例正常对照患者的浆肌活检组织，培养肠成纤维细胞。用Western印迹和定量逆转录聚合酶链式反应(qRT-PCR)检测N-钙粘蛋白的表达。用转化生长因子-β(1)刺激成纤维细胞，用选择性信号通路抑制剂Y27632、PD98050和LY294002分别检测Rho/ROCK、ERK-1/2和Akt信号通路。细胞迁移采用划痕实验进行评估。结果：与直接邻近的正常肠成纤维细胞相比，纤维狭窄CD成纤维细胞表达的N-钙粘蛋白基因和蛋白增加，基底细胞迁移增强。经转化生长因子-β(1)处理的对照成纤维细胞以剂量依赖的方式诱导N-钙粘蛋白，而Rho/ROCK和Akt信号通路的调节抑制了该作用。对照组成纤维细胞在经转化生长因子-β(1)或N-钙粘蛋白载体转染后细胞迁移增强。结论：CD狭窄处成纤维细胞表达结构性N-钙粘附素，并表现为基底细胞迁移增强。转化生长因子-β(1)是肠成纤维细胞中N-钙粘附素的有效诱导剂，可促进细胞迁移。转化生长因子-β(1)介导的N-钙粘附素的诱导可能会加强Crohn‘s狭窄的形成。
Background: Intestinal fibroblasts mediate stricture formation in Crohn's disease (CD). Transforming growth factor-beta(1) (TGF-beta(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 = 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-beta(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-beta(1) 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-beta(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-beta(1) is a potent inducer of N-cadherin in intestinal fibroblasts resulting in enhanced cell migration. The TGF-beta(1)-mediated induction of N-cadherin may potentiate Crohn's stricture formation.