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Regulatory mechanism to tissue factor gene expression by its methylation/demethylation

组织因子基因甲基化/去甲基化的调控机制

基本信息

批准号：
09671108
负责人：
NAKAMURA Shin
金额：
$ 2.18万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

项目摘要

Tissue Factor (TF) is a factor VII/VIIa receptor that initiates blood coagulation. TF is suggested to have other functions such as involvement in embryogenesis, angiogenesis, tumor metastasis, and/or signal transdution. TF gene expression is regulated by several is-elements of AP-1, kappaB, SP-1 and Erg-1 and transcriptional factors to bind these cis-elements. Another regulatory machinery to TF gene expression is expected because there are a lot of CG island specific site for methylation at cytosine residues.In this studies we examined a possible regulation mechanism of TF gene expression through methylation and/or demethylation of its promoter region. We also investigated a relationship between differentiation and de-differentiation/transformation of several cells and methylation/ demethylation of TF gene.We established a competitive RT-PCR which is enable to quantitative measurement of TF m-RNA expressed in several normal or transformed cells using specific primer sets. We also established sensitive sandwich enzyme-linked immuno-sorbent assay to measure TF antigen produced in these cells.In normal blood cells monocytes and granulocytes expressed TF but lymphocytes did not. Marked different expression of TF gene in monkey male reproductive tissues. Seminal vesicle specifically expressed very high level of TF-mRNA.These suggest TF gene expression should regulated by methylation/demethylation during differentiation of leukocytes from common precursor and tissue speculation.

组织因子(Tf)是一种启动凝血的凝血因子VII/VIIa受体。Tf还具有其他功能，如参与胚胎发生、血管生成、肿瘤转移和/或信号传导。TF基因的表达受AP-1、kappaB、SP-1和Erg-1等IS元件以及结合这些顺式元件的转录因子的调控。由于胞嘧啶残基上存在大量的CG岛特异性甲基化位点，另一种对TF基因表达的调控机制被认为是可能的。在本研究中，我们研究了通过其启动子区域的甲基化和/或去甲基化来调控TF基因表达的可能机制。我们还研究了几种细胞分化和去分化/转化与转铁蛋白基因甲基化/去甲基化的关系。我们建立了竞争性RT-PCR方法，可以利用特定的引物组来定量检测几种正常或转化细胞中转铁蛋白m-RNA的表达。我们还建立了敏感的夹心酶联免疫吸附试验来检测这些细胞产生的TF抗原。在正常血细胞中，单核细胞和粒细胞表达TF，而淋巴细胞不表达。Tf基因在猕猴雄性生殖组织中的表达差异显著。精囊特异性表达很高水平的Tf-mRNA，提示在白细胞分化和组织推测过程中，Tf基因的表达可能受到甲基化/去甲基化的调节。