Detection of up-regulated genes in thrombin-stimulated human umbilical vein endothelial cells |
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Authors: | Okada Makoto Suzuki Kimihiro Takada Kunio Nakashima Masahiro Nakanishi Takashi Shinohara Tadashi |
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Affiliation: | aInternal Medicine I, National Defense Medical College, Namiki 3-2, Tokorozawa, Saitama, 359-8513, Japan bDivision of Molecular Neurobiology, Department of Basic Medical Sciences, Institute of Medical Science, University of Tokyo, Tokyo, Japan |
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Abstract: | INTRODUCTION: Thrombin, a serine protease, plays an important role in such actions as coagulation, cell proliferation and inflammation. It has been sporadically reported that endothelial cells, when stimulated by thrombin via protease-activated receptors (PAR), express various mediators and proteins including cytokines, chemokines, growth factors, and adhesion molecules. However, the pleiotropic effect of thrombin on endothelial cells has not yet been fully elucidated. MATERIALS AND METHODS: We newly searched for the up-regulated genes in the thrombin-stimulated endothelial cells by thorough screening using a microarray chip, printed with 22,575 human genes, followed by verification using real-time PCR (n=3). RESULTS AND CONCLUSIONS: Twelve genes, which were 4.8 times or more up-regulated in a microarray analysis, were selected and further analyzed. In real-time PCR, ICAM-1, IL-8, BIRC3, COL3A1, CXCL3, and CXCL1 were significantly up-regulated in the thrombin-stimulated cells: 16.0-, 8.81-, 5.92-, 3.74-, 1.74-, and 1.66-fold, respectively. VCAM-1, CXCL2, CCL20, CSF2, CD69, and CCL2 were up-regulated in the thrombin-stimulated cells: 12.2-, 2.44-, 1.90-, 1.82-, 1.62-, and 1.06-fold, respectively, without attaining statistical significance. We demonstrated, for the first time, that BIRC3 (anti-apoptotic protein), COL3A1 (matrix protein synthesis), and CXCL3 (chemokine) were up-regulated in the thrombin-stimulated HUVECs. |
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Keywords: | Thrombin Human umbilical vein endothelial cell Gene expression Microarray Real-time PCR |
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