© 1999 American Heart Association, Inc.
Original Contribution |
Vascular Endothelial Growth Factor–Induced Endothelial Cell Migration and Proliferation Depend on a Nitric Oxide–Mediated Decrease in Protein Kinase C
Activity
From the Cardiovascular Division, Department of Medicine (Y.S., S.T., R.Y., J.A.W.), and the Department of Molecular Pharmacology (J.A.W.), Albert Einstein College of Medicine/Montefiore Medical Center, Bronx, NY.
Correspondence to J. Anthony Ware, MD, Albert Einstein College of Medicine, Forchheimer Bldg, G-46, 1300 Morris Park Ave, Bronx, NY 10461. E-mail jaware{at}aecom.yu.edu
Abstract—Vascular
endothelial growth factor (VEGF) promotes angiogenesis
and endothelial cell (EC) migration and proliferation
by affecting intracellular mediators, only some of which are known,
distal to its receptors. Protein kinase C (PKC) participates in the
function of VEGF, but the role of individual PKC isoenzymes is unknown.
In this study, we tested the importance of the activity of specific PKC
isoenzymes in human EC migration and proliferation in response to VEGF.
PKC
specific activity was depressed by the addition of VEGF (by
41±8% [P<0.05] at 24 hours) in human umbilical vein ECs (HUVECs)
and in a HUVEC-derived EC line, ECV, without changing the total amount
of either protein or mRNA encoding PKC. Neither basic fibroblast
growth factor (FGF-2) nor serum altered PKC specific
activity. The VEGF-induced decrease of PKC activity, which began at
8 hours after stimulation, was strongly blocked by pretreatment with
the nitric oxide (NO) synthase inhibitor
NG-monomethyl-L-arginine
in HUVECs; NO release peaked within 2 hours after stimulation. An
exogenous NO donor, sodium nitroprusside, also decreased PKC
activity. The inhibition by
NG-monomethyl-L-arginine
of VEGF-induced HUVEC migration and proliferation, but not that induced
by FGF-2 or serum, suggested that the decrease in PKC via NO pathway
is required for VEGF-induced EC migration and proliferation.
Overexpression of PKC in ECV cells specifically prevented EC
response to VEGF but not to FGF-2 or serum. Thus, we conclude that
suppression of PKC activity via a NO synthase mechanism is required
for VEGF-induced EC migration and proliferation, but not for that
induced by FGF-2 or serum.
Key Words: endothelium • growth factor • nitric oxide synthase • intracellular signaling • angiogenesis
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