Gene expression analysis of HUVEC in response to TF-binding
Gene expression analysis of HUVEC in response to TF-binding
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DOI：
10.1016/j.thromres.2010.11.024
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发表时间：
2011-03-01
影响因子：
7.5
通讯作者：
Albrecht, Sybille
中科院分区：
文献类型：
--
作者：
Grosser, Marianne;Magdolen, Viktor;Albrecht, Sybille
简介：组织因子（TF）是因子VII/VIIa（FVII/FVIIa）的辅因子和外源性途径的起始因子，在细胞因子和生长因子的控制下在血管内细胞上瞬时表达。此外，内皮细胞表达外部TF的结合位点。在本研究中，我们研究了来自人脐静脉内皮细胞（HUVEC）的基因表达，在响应TF-结合，以确定差异表达的gene.Materials和Methods：HUVEC与重组relipidated TF（Innovin）与未处理的细胞，以及TF/FVIIa与FVIIa单独处理。通过ELISA测量TF结合。结果：TF作用后4 h，HUVEC基因表达谱中有148条上调，29条下调。值得注意的是，无论是通过单独的TF还是通过TF/FVIIa的复合物，显著上调和下调的基因都表现出完全重叠，表明在结合外部添加的TF后内皮细胞的活化不依赖于FVIIa，如TF表达细胞所证明的那样。通过qPCR证实TF介导的几种基因的基因表达调节，涉及细胞凋亡、细胞粘附、细胞运动和血管生成的调节。此外，在SELE的情况下，TGFB 2，TNFAIP 3，TNFSF 4，TNFSF 18，TAGLN，CXCL 1，PCF 11抗体直接TF明显抑制TF介导的调节基因expression.Conclusions：结果表明，TF与HUVEC通过一个结合位点的相互作用，独立于FVIIa，可能会导致调节各种基因参与动脉硬化，癌症和心血管疾病。(C)2010爱思唯尔有限公司版权所有。
Introduction: Tissue factor (TF), the cofactor for factor VII/VIIa (FVII/FVIIa) and initiator of the extrinsic pathway, is transiently expressed on intravascular cells under control of cytokines and growth factors. In addition, endothelial cells express a binding site for external TF. In the present study, we investigated gene expression of endothelial cells derived from human umbilical veins (HUVEC) in response to TF-binding to identify differentially expressed genes.Materials and methods: HUVEC were treated with recombinant relipidated TF (Innovin) versus nontreated cells, as well as TF/FVIIa versus FVIIa alone. TF binding was measured by ELISA. Gene expression profiles were examined using HG-U133 plus 2.0 arrays (Affymetrix).Results: Gene expression analysis of HUVEC showed 148 up-regulated and 29 down-regulated genes 4 h after TF binding. Notably, the genes, which were significantly up-and down-regulated, either by TF alone or by the complex of TF/FVIIa, exhibited a complete overlap, indicating that activation of endothelial cells after binding of external added TF does not depend on FVIIa as has been demonstrated for TF-expressing cells. TF-mediated regulation of gene expression of several genes, involved in regulation of apoptosis, cell adhesion, cell motility, and angiogenesis, was confirmed by qPCR. Furthermore, in case of SELE, TGFB2, TNFAIP3, TNFSF4, TNFSF18, TAGLN, CXCL1, PCF11 antibodies directed to TF clearly inhibited TF-mediated regulation of gene expression.Conclusions: The results demonstrate that interaction of TF with HUVEC via a binding site, independent from FVIIa, may result in regulation of a variety of genes involved in arteriosclerosis, cancer, and cardiovascular diseases. (C) 2010 Elsevier Ltd. All rights reserved.