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Cloning of shear stress-sensitive endothelial genes

剪切应力敏感内皮基因的克隆

基本信息

批准号：
08408039
负责人：
ANDO Joji
金额：
$ 13.76万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08408039/
关键词：
shear stress endothelial cell mRNA differential display G-protein gene 内皮細胞壁ずり応力血行力学因子

项目摘要

The purpose of this study was to investigate endothelial gene which were responsive to shear stress generated by blood flow and to find genes encoding new bio-active substances. First, in order to evaluate whole number of shear stress-responsive genes in endothelial cells, differential display of mRNAs was performed. The results showed that 33 of 15O7mRNAs detected were up-regulated by a shear stress of 15 dynes/cm^2 for 6 h, while 27 were down-regulated, indicating that around 4% of the whole endothelial genes are shear-sensitive. Second, 16 genes were cloned from these 60 shear-sensitive genes and sequenced. Homology search revealed 6 known genes encoding laminin B1 chain, H^+-ATP synthase coupling factor 6, lysyl oxidase, myosin light chain kinase, NADH dehydrogenase, and interleukin-8 receptor and 10 unknown genes. Lastly, we constructed a cDNA library from human umbilical vein endothelial cells which had been exposed to a shear stress of 15 dynes/cm^2 for 8 h, and looked for new G-protein coupled receptors. As a result we obtained a new one which we named PEG-1 (Flow-induced Endothelial G-protein-coupled receptor. The receptor consists of 375 amino acids and has a homology to angiotensin receptors or chemokine receptors.

本研究的目的是研究血管内皮细胞对血流产生的剪切力的反应基因，并寻找编码新的生物活性物质的基因。首先，为了评估内皮细胞中剪切力反应基因的总数，进行了mRNAs的差异显示。结果显示，15O7mRNAs在15dynes/cm2的剪切力作用6h后，33个上调，27个下调，表明约4%的内皮细胞基因是剪切敏感的。其次，从这60个剪切敏感基因中克隆了16个基因并进行了测序。同源性检索发现了6个编码层粘连蛋白B1链、H^+-ATP合成酶偶联因子6、赖氨酰氧化酶、肌球蛋白轻链激酶、NADH脱氢酶和白细胞介素8受体的已知基因和10个未知基因。最后，我们从暴露于15dynes/cm2剪切力8h的人脐静脉内皮细胞中构建了一个cDNA文库，并寻找新的G蛋白偶联受体。因此，我们获得了一种新的受体，我们将其命名为PEG1(Flow-Induced EndothelialG-Protein-Couted Receptor，流诱导内皮G蛋白偶联受体)。该受体由375个氨基酸组成，与血管紧张素受体或趋化因子受体具有同源性。