Abstract
Vascular endothelial growth factor-A (VEGF), which binds to both VEGF receptor-1 (Flt1) and VEGFR-2 (KDR/Flk-1), requires nitric oxide (NO) to induce angiogenesis in a cGMP-dependent manner. Here we show that VEGF-E, a VEGFR-2-selective ligand stimulates NO release and tube formation in human umbilical vein endothelial cells (HUVEC). Inhibition of phospholipase Cgamma (PLCgamma) with U73122 abrogated VEGF-E induced endothelial cell migration, tube formation and NO release. Inhibition of endothelial nitric oxide synthase (eNOS) using l-NNA blocked VEGF-E-induced NO release and angiogenesis. Pre-incubation of HUVEC with the soluble guanylate cyclase inhibitor, ODQ, or the protein kinase G (PKG) inhibitor, KT-5823, had no effect on angiogenesis suggesting that the action of VEGF-E is cGMP-independent. Our data provide the first demonstration that VEGFR-2-mediated NO signaling and subsequent angiogenesis is through a mechanism that is dependent on PLCgamma but independent of cGMP and PKG.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Carbazoles / pharmacology
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Cell Movement / drug effects
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Cells, Cultured
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Cyclic GMP / metabolism*
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Cyclic GMP-Dependent Protein Kinases / antagonists & inhibitors
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Cyclic GMP-Dependent Protein Kinases / metabolism*
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Dose-Response Relationship, Drug
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Endothelial Cells / cytology
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Endothelial Cells / drug effects*
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Endothelial Cells / metabolism
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Enzyme Activation / drug effects
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Enzyme Inhibitors / pharmacology
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Estrenes / pharmacology
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Humans
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Indoles / pharmacology
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Neovascularization, Physiologic / drug effects
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Nitric Oxide / metabolism
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Nitric Oxide Synthase / antagonists & inhibitors
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Nitric Oxide Synthase / metabolism*
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Nitroarginine / pharmacology
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Phospholipase C gamma / antagonists & inhibitors
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Phospholipase C gamma / metabolism
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Phosphorylation / drug effects
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Pyrrolidinones / pharmacology
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Signal Transduction / drug effects*
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Swine
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Vascular Endothelial Growth Factor Receptor-2 / metabolism
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Viral Proteins / pharmacology*
Substances
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Carbazoles
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Enzyme Inhibitors
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Estrenes
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Indoles
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Pyrrolidinones
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VEGF-like protein, Orf virus
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Viral Proteins
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1-(6-((3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione
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KT 5823
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Nitroarginine
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Nitric Oxide
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Nitric Oxide Synthase
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Vascular Endothelial Growth Factor Receptor-2
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Cyclic GMP-Dependent Protein Kinases
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Phospholipase C gamma
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Cyclic GMP