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J. Biol. Chem., Vol. 276, Issue 15, 11830-11837, April 13, 2001
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v
3-
and 1-containing Integrins*
From the Center for Vascular Biology, Department of Physiology,
University of Connecticut Health Center,
Farmington, Connecticut 06030-3501
Sphingosine 1-phosphate (SPP), a platelet-derived
bioactive lysophospholipid, is a regulator of angiogenesis. However,
molecular mechanisms involved in SPP-induced angiogenic responses are
not fully defined. Here we report the molecular mechanisms involved in
SPP-induced human umbilical vein endothelial cell (HUVEC) adhesion and
migration. SPP-induced HUVEC migration is potently inhibited by
antisense phosphothioate oligonucleotides against EDG-1 as well as
EDG-3 receptors. In addition, C3 exotoxin blocked SPP-induced cell
attachment, spreading and migration on fibronectin-, vitronectin- and
Matrigel-coated surfaces, suggesting that endothelial differentiation gene receptor signaling via the Rho pathway is critical for SPP-induced cell migration. Indeed, SPP induced Rho activation in an
adherence-independent manner, whereas Rac activation was dispensible
for cell attachment and focal contact formation. Interestingly, both
EDG-1 and -3 receptors were required for Rho activation. Since
integrins are critical for cell adhesion, migration, and angiogenesis,
we examined the effects of blocking antibodies against
v
3, 1, or
3 integrins. SPP induced Rho-dependent
integrin clustering into focal contact sites, which was essential for
cell adhesion, spreading and migration. Blockage of
v3- or 1-containing
integrins inhibited SPP-induced HUVEC migration. Together our results
suggest that endothelial differentiation gene receptor-mediated Rho
signaling is required for the activation of integrin
v3 as well as 1-containing
integrins, leading to the formation of initial focal contacts and
endothelial cell migration.
To whom correspondence should be addressed: Center for Vascular
Biology, Dept. of Physiology, University of Connecticut Health Center,
263 Farmington Ave., Farmington, CT 06030-3501. Tel.: 860-679-4128;
Fax: 860-679-1201; E-mail: hla@sun.uchc.edu.
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