Expression Profiling of Wnt Family of Genes in Normal and Inflammatory Bowel Disease Primary Human Intestinal Myofibroblasts and Normal Human Colonic Crypt Epithelial Cells
Expression Profiling of Wnt Family of Genes in Normal and Inflammatory Bowel Disease Primary Human Intestinal Myofibroblasts and Normal Human Colonic Crypt Epithelial Cells
复制标题
DOI：
10.1002/ibd.21353
复制
发表时间：
2011-01-01
影响因子：
4.9
通讯作者：
Mahida, Y. R.
中科院分区：
文献类型：
--
作者：
Hughes, K. R.;Sablitzky, F.;Mahida, Y. R.
背景：Wnt信号调节肠上皮干细胞的功能。Wnt配体结合卷曲（Fz）受体和低密度脂蛋白受体相关蛋白（LRP） 5和6。分泌卷曲相关蛋白（SFRP）和Dickkopf家族抑制Wnt信号传导。我们的目的是研究Wnt家族基因在分离的肠肌成纤维细胞和隐窝上皮细胞中的表达。方法：从正常结肠和小肠粘膜、溃疡性结肠炎（UC）和克罗恩病（Crohn's disease）患者中分离肌成纤维细胞。采用聚合酶链反应（PCR）技术检测Wnt家族基因的表达。用IEC-6细胞研究上皮细胞增殖。结果：大多数分离的肌成纤维细胞表达Wnt2、Wnt5A、Wnt5B、Fzd1、Fzd2、Fzd4、Fzd6、Fzd7、Fzd8、LRP6、Dick-kopf1和SFRP1。与从正常结肠粘膜样本中分离的肌成纤维细胞相比，实时逆转录- pcr研究（使用其他分离物）显示UC肌成纤维细胞中SFRP1的表达显著降低（降低3.34倍，P < 0.01）。重组SFRP1抑制IEC-6上皮细胞的增殖。在结肠隐窝上皮细胞中，Wnt配体及其抑制剂的表达通常不存在或非常弱。相比之下，所有隐窝上皮制剂均表达Fzd1、Fzd5、Fzd7、Fzd8和LRP6。结论：人肠肌成纤维细胞表达了许多Wnt配体、它们的受体和抑制剂。相反，结肠隐窝上皮细胞主要表达Wnt受体。与从正常结肠粘膜分离的肌成纤维细胞相比，UC患者的SFRP1表达显著降低。由于SFRP1表达减少与恶性肿瘤有关，因此这种Wnt抑制剂的肌成纤维细胞低表达可能与UC中癌症风险增加有关。
Background: Wnt signaling regulates intestinal epithelial stem cell function. Wnt ligands bind Frizzled (Fz) receptors and low-density lipoprotein-receptor-related protein (LRP) 5 and 6. Secreted Frizzled-related protein (SFRP) and Dickkopf families inhibit Wnt signaling. Our aim was to study expression of Wnt family of genes in isolated intestinal myofibroblasts and crypt epithelial cells.Methods: Myofibroblasts were isolated from normal colonic and small intestinal mucosal samples and those affected by ulcerative colitis (UC) and Crohn's disease. Expression of the Wnt family of genes was studied by polymerase chain reaction (PCR) array. Epithelial proliferation was studied using IEC-6 cells.Results: Most of the myofibroblast isolates expressed Wnt2, Wnt5A, Wnt5B, Fzd1, Fzd2, Fzd4, Fzd6, Fzd7, Fzd8, LRP6, Dick-kopf1, and SFRP1. Compared to myofibroblasts isolated from normal colonic mucosal samples, real-time reverse transcription-PCR studies (using additional isolates) showed significantly reduced expression of SFRP1 in UC myofibroblasts (3.34-fold reduction, P < 0.01). Recombinant SFRP1 inhibited proliferation of IEC-6 epithelial cells. In colonic crypt epithelial cells, expression of Wnt ligands and their inhibitors was generally either absent or very weak. By contrast, all the crypt epithelial preparations expressed Fzd1, Fzd5, Fzd7, Fzd8, and LRP6.Conclusions: Human intestinal myofibroblasts expressed a number of Wnt ligands, their receptors, and inhibitors. In contrast, colonic crypt epithelial cells predominantly expressed Wnt receptors. Compared to myofibroblasts isolated from normal colonic mucosa, those affected by UC showed significantly reduced expression of SFRP1. Since reduced SFRP1 expression has been associated with malignancy, low myofibroblast expression of this Wnt inhibitor may be implicated in increased risk of cancer in UC.