Fibroblast-derived HB-EGF promotes Cdx2 expression in esophageal squamous cells
Fibroblast-derived HB-EGF promotes Cdx2 expression in esophageal squamous cells
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DOI：
10.1038/labinvest.2010.71
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发表时间：
2010-07-01
影响因子：
5
通讯作者：
Kinoshita, Yoshikazu
中科院分区：
文献类型：
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作者：
Rahman, Farzana B.;Kadowaki, Yasunori;Kinoshita, Yoshikazu
Barrett食管柱状表型的分子基础尚不清楚。一种假说认为，慢性反流性食管炎中由间质起源细胞局部产生的因子可诱导化生转化。本研究旨在阐明纤维母细胞分泌的因子导致邻近鳞状上皮的柱状表型。将人成纤维细胞置于酸化培养基中20 min，培养基中和2 h，然后将总RNA杂交到Sentrix Human-6 Expression BeadChips上。此外，应用免疫组织化学方法检测反流性食管炎患者食管黏膜活检标本中HB-EGF的表达。此外，用重组HB-EGF处理人食管鳞状上皮细胞系HET1A细胞，分析Cdx2和柱状标记物的表达变化。基因表达谱显示，在酸暴露的反应中，多种生长因子和炎症趋化因子显著上调。其中HB-EGF上调10倍以上。来自反流性食管炎患者的活检标本显示，受损上皮下的成纤维细胞中HB-EGF表达强烈。此外，HB-EGF对HET1A细胞的体外刺激使Cdx2呈剂量依赖性增加。对人Cdx2启动子的功能分析也揭示了其在HB-EGF刺激下的上调，显示了潜在响应元件（AP-1和NF-kappa B）在其转录激活中的作用。此外，柱状标志物细胞角蛋白7和绒毛蛋白也在HB-EGF刺激下上调。HB-EGF以旁分泌方式诱导食管鳞状上皮柱状表型的几个基因特征。实验室调查（2010）90,1033-1048；doi: 10.1038 / labinvest.2010.71;2010年3月29日在线发布
The molecular basis of attaining columnar phenotype in Barrett's esophagus is poorly understood. One hypothesis states that factors locally produced by cells of mesenchymal origin in chronic reflux esophagitis induce metaplastic transformation. This study was performed to elucidate the factors secreted from fibroblasts that cause columnar phenotype in adjacent squamous epithelium. Human fibroblast cells were exposed to acidified medium for 20 min, followed by medium neutralization for 2 h, and then total RNA was hybridized to Sentrix Human-6 Expression BeadChips. Furthermore, esophageal mucosal biopsy specimens from reflux esophagitis patients were examined for HB-EGF expression using immunohistochemistry. In addition, cells from the human esophageal squamous epithelial cell line HET1A were treated with recombinant HB-EGF, and changes in expressions of Cdx2 and columnar markers were analyzed. The gene expression profile revealed significant upregulation of a variety of growth factors and inflammatory chemokines in response to acid exposure. Among them, HB-EGF was upregulated more than 10-fold. Biopsy specimens from reflux esophagitis patients showed a strong expression of HB-EGF in fibroblast cells underlying the damaged epithelium. Furthermore, in vitro stimulation of HET1A cells with HB-EGF increased Cdx2 in dose-dependent manners. Functional analysis of human Cdx2 promoter also revealed its upregulation by HB-EGF stimulation, showing the role of potential responsive elements (AP-1 and NF-kappa B) for its transcriptional activation. Moreover, the columnar markers cytokeratin 7 and villin were also upregulated by HB-EGF stimulation. HB-EGF induces several genes characteristics of columnar phenotypes of esophageal squamous epithelium in a paracrine manner. Laboratory Investigation (2010) 90, 1033-1048; doi:10.1038/labinvest.2010.71; published online 29 March 2010