Detection of up-regulated genes in thrombin-stimulated human umbilical vein endothelial cells.
BACKGROUND
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.


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).


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.
Okada
Makoto
M
Internal Medicine I, National Defense Medical College, Namiki 3-2, Tokorozawa, Saitama, 359-8513, Japan. grd1804@ndmc.ac.jp
Suzuki
Kimihiro
K
Takada
Kunio
K
Nakashima
Masahiro
M
Nakanishi
Takashi
T
Shinohara
Tadashi
T
D000075747:Baculoviral IAP Repeat-Containing 3 Protein; D018929:Cell Culture Techniques; D019743:Chemokines, CXC / Q000378:metabolism*; D024061:Collagen Type III / Q000378:metabolism*; D042783:Endothelial Cells / Q000378:metabolism; D020869:Gene Expression Profiling; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D013917:Thrombin / Q000502:physiology*; D044767:Ubiquitin-Protein Ligases; D014471:Umbilical Veins / Q000378:metabolism; D015854:Up-Regulation
C439971:COL3A1 protein, human; C499707:CXCL3 protein, human; D019743:Chemokines, CXC; D024061:Collagen Type III; D051034:Inhibitor of Apoptosis Proteins; C494832:BIRC3 protein, human; D000075747:Baculoviral IAP Repeat-Containing 3 Protein; D044767:Ubiquitin-Protein Ligases; D013917:Thrombin