TREFOIL FACTORS AND ORAL MUCOSAL WOUND HEALING

三叶因子是一个由三个小肽组成的家族，它们与粘蛋白一起分泌到肠腔内，在肠上皮损伤后上皮完整性的恢复中发挥关键作用。我们建议检验这一假设，即一个或多个三叶因子也有助于 维持口腔内上皮的完整性。 该项目的具体目标是： 1.采用RT-PCR和原位杂交相结合的方法，确定口腔黏膜上皮细胞是否产生三叶因子(PS2/TFF1、SP/TFF2和ITF/TFF3)中的任何一种。 2.建立一种标准化的口腔上皮细胞迁移和损伤修复的体外检测方法，以确定口腔上皮细胞对重组肠三叶因子(ITF/TFF3)的反应是否伴随着其运动特性的改变。 3.使用融合蛋白分析口腔上皮细胞系是否存在能够与ITF/TFF3结合的表面受体，该融合蛋白包括用于位点特异性酶生物素化的共同序列。 寻找更多促进口腔黏膜溃烂病变愈合的天然多肽是开发新的治疗方法的第一步，这些新的治疗方法是治疗复发性严重口腔溃疡和化疗引起的粘膜炎等重要临床问题的第一步。

Trefoil factors are a family of three small peptides secreted into the gut lumen with mucins that play a critical role in restoration of epithelial integrity following injury to gut epithelium. We propose to test the hypothesis that one or more of the trefoil factors also contribute to maintenance of epithelial integrity in the oral cavity. The specific aims of this project are: 1. To determine whether epithelial cells lining the oral cavity make any of the 3 defined human trefoil factors (pS2/TFF1, SP/TFF2, and ITF/TFF3) using a combination of RT-PCR and in situ hybridization. 2. To determine whether oral cavity epithelial cell lines can respond to recombinant intestinal trefoil factor (ITF/TFF3) with changes in their motogenic properties in a well-standardized in vitro assay for cell migration and epithelial wound healing. 3. To analyze oral cavity epithelial cell lines for the presence of, surface receptors capable of binding to ITF/TFF3 using a fusion protein that includes a consensus sequence for site-specific enzymatic biotinylation. Identification of additional natural polypeptides that favor healing of ulcerative lesions on oral mucosa is a first step towards development of new therapeutic approaches to the important clinical problems of recurrent severe apthous ulcers and chemotherapy-induced mucositis.