Expression profiling of ETS and MMP factors in VEGF-activated endothelial cells: role of MMP-10 in VEGF-induced angiogenesis.
In the process of angiogenesis, working of many transcription factors at the proper time is important to activate angiogenesis-related genes such as cytokine, matrix protease and adhesion molecules. In this study, we searched for Ets transcription factors and matrix metalloproteinases (MMPs) that respond to VEGF in endothelial cells. We first analyzed the expression of 27 human Ets factors and 15 human MMPs in VEGF-treated human umbilical vein endothelial cells (HUVEC) using quantitative RT-PCR. The most abundant Ets factors in HUVEC were ETS-1, Fli-1, ERP/NET/ELK3, and ERG. MMP-1, -2, -10, -11, -14, -15, and -16 were also detected in HUVEC. We also found that ETV-1, Fli-1, ERG, MMP-1, -3, -7, -8, -9, -10, -13, and -19 expression is up-regulated more than 1.5-fold in HUVEC after 2 h of VEGF treatment. In addition, the expression of MMP-10 induced by VEGF remained twofold higher for 24 h compared to non-treated control. The elevation of MMP10 mRNA and protein levels was confirmed to be both time- and dosage-dependent. In addition, MMP-10 transcription was mediated by Ets-1 but not ERP/NET/ELK3. The inhibition of PI3K and MAPK inhibited VEGF-induced MMP-10 expression. Furthermore, transfection of MMP-10 siRNA inhibited VEGF-induced migration and tube formation in HUVEC, and it also inhibited vessel formation in matrigel plugs in vivo. In conclusion, our study demonstrated induction of MMP-10 by VEGF in HUVEC and supports an angiogenic role for MMP-10 in response to VEGF stimulation in vitro and in vivo.
Heo
Sun-Hee
SH
Department of Biochemistry, School of Dentistry, Kyungpook National University, Daegu, Korea.
Choi
Young-Jin
YJ
Ryoo
Hyun-Mo
HM
Cho
Je-Yoel
JY
D043925:Angiogenesis Inducing Agents / Q000494:pharmacology*; D000818:Animals; D002460:Cell Line; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects* / Q000502:physiology*; D004730:Endothelium, Vascular / Q000166:cytology; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D053506:Matrix Metalloproteinase 10 / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D008969:Molecular Sequence Data; D018919:Neovascularization, Physiologic; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D050783:Proto-Oncogene Proteins c-ets / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D015398:Signal Transduction / Q000502:physiology; D042442:Vascular Endothelial Growth Factors / Q000494:pharmacology*
D043925:Angiogenesis Inducing Agents; D007527:Isoenzymes; D050783:Proto-Oncogene Proteins c-ets; D042442:Vascular Endothelial Growth Factors; D019869:Phosphatidylinositol 3-Kinases; D020928:Mitogen-Activated Protein Kinases; D053506:Matrix Metalloproteinase 10