J Korean Med Sci.  2002 Apr;17(2):161-167. 10.3346/jkms.2002.17.2.161.

Expression and Regulation of Endothelial Nitric Oxide Synthase by Vascular Endothelial Growth Factor in ECV 304 Cells

• Park JS
• Hong GR
• Baek SW
• Shin DG
• Kim YJ
• Shim BS

• KMID: 648670
• DOI: http://doi.org/10.3346/jkms.2002.17.2.161

Abstract

Nitric oxide (NO) seems to play a pivotal role in the vascular endothelial growth factor (VEGF)-induced endothelial cell proliferation. This study was designed to investigate the role and intracellular signal pathway of endothelial nitric oxide synthase (eNOS) activation induced by VEGF. ECV 304 cells were treated with betaVEGF(165) and then cell proliferation, eNOS protein and mRNA expression levels were analyzed to elucidate the functional role of eNOS in cell proliferation induced by VEGF. After exposure of cells to betaVEGF(165) , eNOS activity and cell growth were increased by approximately two-fold in the betaVEGF(165) -treated cells compared to the untreated cells. In addition, VEGF stimulated eNOS expression at both the mRNA and protein levels in a dose-dependent manner. Phosphatidylinositol-3 kinase (PI-3K) inhibitors were used to assess PI-3K involvement in eNOS regulation. LY294002 was found to attenuate VEGF-stimulated eNOS expression. Wortmannin was not as effective as LY294002, but the reduction effect was detectable. Cells activated by VEGF showed increased ERK1/2 levels. Moreover, the VEGF-induced eNOS expression was reduced by the PD98059, MAPK pathway inhibitor. This suggests that eNOS expression might be regulated by PI-3K and the ERK1/2 signaling pathway. In conclusion, betaVEGF(165) induces ECV 304 cell proliferation via the NO produced by eNOS. In addition, eNOS may be regulated by the PI-3K or mitogen-activated protein kinase pathway.

Keyword

Angiogenesis; Nitric-Oxide Synthase

MeSH Terms

1-Phosphatidylinositol 3-Kinase/*antagonists & inhibitors
Cell Division/drug effects
Cell Line
Endothelial Growth Factors/*metabolism/pharmacology
Endothelium, Vascular/cytology
*Gene Expression Regulation, Enzymologic
Lymphokines/*metabolism/pharmacology
MAP Kinase Signaling System
Mitogen-Activated Protein Kinase 1/*antagonists & inhibitors
Mitogen-Activated Protein Kinase 3
Mitogen-Activated Protein Kinases/*antagonists & inhibitors
Nitric Oxide Synthase/*genetics/metabolism
Nitric Oxide Synthase Type III
Signal Transduction
Vascular Endothelial Growth Factor A
Vascular Endothelial Growth Factors