Differential regulation of sphingosine-1-phosphate- and VEGF-induced endothelial cell chemotaxis involvement of Giα2-linked Rho kinase activity

F. Liu, A. D. Verin, P. Wang, R. Day, R. P. Wersto, F. J. Chrest, D. K. English, J. G.N. Garcia

Research output: Contribution to journalArticlepeer-review

138 Scopus citations


We compared stimulus-coupling pathways involved in bovine pulmonary artery (PA) and lung microvascular endothelial cell migration evoked by sphingosine-1-phosphate (S1P), a potent bioactive lipid released from activated platelets, and by vascular endothelial growth factor (VEGF), a well-recognized angiogenic factor. S1P-induced endothelial cell migration was maximum at 1 μM (∼ 8-fold increase with PA endothelium) and surpassed the maximal response evoked by either VEGF (10 ng/ml) (∼ 2.5-fold increase) or hepatocyte growth factor (HGF) (∼ 2.5-fold increase). Migration induced by S1P, but not by VEGF, was significantly inhibited by treatment with antisense oligonucleotides directed to Edg-1 and Edg-3 (endothelial differentiation gene) S1P receptors and by G protein modification. These strategies included pretreatment with pertussis toxin, or transfection with mini-genes encoding a βγ subunit inhibitory peptide of the β-adrenergic receptor kinase, or an 11-amino-acid peptide that inhibits G1α2 signaling. Various strategies to interrupt Rho family signaling, including C3 exotoxin, dominant/negative Rho, or the addition of Y27632, a cell-permeable Rho kinase inhibitor, significantly attenuated S1P- but not VEGF-induced migration. Conversely, pharmacologic inhibition of either myosin light chain kinase, src family tyrosine kinases, or phosphatidylinositol-3′ kinase reduced basal endothelial cell migration and abolished VEGF-induced endothelial cell migration but did not inhibit the increase in S1P-induced migration. Whereas VEGF and S1P increased both p42/p44 extracellular regulated kinase and p38 mitogen-activated protein (MAP) kinase activities, only p38 MAP kinase inhibition significantly reduced VEGF- and S1P-stimulated migration. These data confirm S1P as a potent endothelial cell chemoattractant through G1α2-coupled Edg receptors linked to Rho-associated kinase and p38 MAP kinase activation. The divergence in signaling pathways evoked by S1P and VEGF suggests complex and agonist-specific regulation of endothelial cell angiogenic responses.

Original languageEnglish (US)
Pages (from-to)711-719
Number of pages9
JournalAmerican journal of respiratory cell and molecular biology
Issue number6
StatePublished - 2001
Externally publishedYes

ASJC Scopus subject areas

  • Molecular Biology
  • Pulmonary and Respiratory Medicine
  • Clinical Biochemistry
  • Cell Biology


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