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人内皮细胞中剪切应力反应性信使核糖核酸的差异显示与克隆

Differential display and cloning of shear stress-responsive messenger RNAs in human endothelial cells.

作者信息

Ando J, Tsuboi H, Korenaga R, Takahashi K, Kosaki K, Isshiki M, Tojo T, Takada Y, Kamiya A

机构信息

Department of Cardiovascular Biomechanics, Faculty of Medicine, University of Tokyo, Japan.

出版信息

Biochem Biophys Res Commun. 1996 Aug 14;225(2):347-51. doi: 10.1006/bbrc.1996.1178.

Abstract

To investigate the effect of shear stress on endothelial gene expression, we performed differential display of mRNAs from cultured human umbilical vein endothelial cells either incubated under static conditions or exposed to shear stress (15 dynes/cm2) for 6 h in a flow-chamber. Around 4% of the total number of mRNAs detected were either up- or down-regulated by shear stress. DNA sequencing of some of these shear stress-responsive mRNAs revealed homology of several clones to known gene sequences and many other clones for unknown genes. Known genes, including those for human laminin B1 chain, H(+)-ATP synthase coupling factor 6, lysyl oxidase, myosin light chain kinase, and interleukin-8 receptor, were upregulated by shear stress, while the gene encoding NADH dehydrogenase was down-regulated. The present results suggest that shear stress can change the expression of numerous genes in endothelial cells, far more than reported to date, and that mRNA differential display is quite useful for cloning known and unknown shear stress-responsive genes.

摘要

为了研究剪切应力对内皮细胞基因表达的影响,我们对培养的人脐静脉内皮细胞的mRNA进行了差异显示分析。这些细胞分别在静态条件下孵育,或在流动腔中暴露于剪切应力(15达因/平方厘米)6小时。在检测到的mRNA总数中,约4%的mRNA受剪切应力上调或下调。对其中一些对剪切应力有反应的mRNA进行DNA测序,结果显示一些克隆与已知基因序列具有同源性,还有许多克隆对应未知基因。已知基因,包括人层粘连蛋白B1链、H(+)-ATP合酶偶联因子6、赖氨酰氧化酶、肌球蛋白轻链激酶和白细胞介素-8受体的基因,受剪切应力上调,而编码NADH脱氢酶的基因则下调。目前的结果表明,剪切应力可改变内皮细胞中众多基因的表达,其数量远超迄今报道的数量,而且mRNA差异显示对于克隆已知和未知的剪切应力反应基因非常有用。

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