Institute of Pathology, Technical University of Dresden, D-01307 Dresden, Germany.
Thromb Res. 2011 Mar;127(3):259-63. doi: 10.1016/j.thromres.2010.11.024. Epub 2010 Dec 24.
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.
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).
Gene expression analysis of HUVEC showed 148 up-regulated and 29 down-regulated genes 4h 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.
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.
组织因子（TF）是因子 VII/VIIa（FVII/FVIIa）的辅助因子，也是外源性途径的启动子，在细胞因子和生长因子的控制下，短暂表达于血管细胞。此外，内皮细胞表达外源性 TF 的结合位点。在本研究中，我们研究了人脐静脉内皮细胞（HUVEC）对 TF 结合的基因表达反应，以鉴定差异表达的基因。
用重组再酯化 TF（Innovin）处理 HUVEC 细胞，作为未处理细胞的对照，同时用 TF/FVIIa 处理细胞，作为单独用 FVIIa 处理细胞的对照。通过 ELISA 法测量 TF 结合。使用 HG-U133 plus 2.0 阵列（Affymetrix）进行基因表达谱分析。
HUVEC 的基因表达分析显示，TF 结合后 4 小时有 148 个上调基因和 29 个下调基因。值得注意的是，无论是单独用 TF 还是用 TF/FVIIa 复合物处理，明显上调和下调的基因完全重叠，这表明外源性添加的 TF 结合后内皮细胞的激活不依赖于 FVIIa，这与表达 TF 的细胞已证明的情况一致。qPCR 证实了 TF 对参与细胞凋亡、细胞黏附、细胞运动和血管生成调节的几个基因的表达具有调节作用。此外，在 SELE、TGFB2、TNFAIP3、TNFSF4、TNFSF18、TAGLN、CXCL1、PCF11 基因中，针对 TF 的抗体明显抑制了 TF 介导的基因表达调节。
结果表明，TF 通过与 HUVEC 的结合位点相互作用，不依赖于 FVIIa，可能导致与动脉粥样硬化、癌症和心血管疾病相关的多种基因的调节。
