Sphingosine 1-phosphate stimulates G(i)- and Rho-mediated vascular endothelial cell spreading and migration.
Sphingosine 1-phosphate (Sph-1-P) is a bioactive lipid released from activated platelets, which may be involved in angiogenesis. We, hence, investigated Sph-1-P effects on human umbilical vein endothelial cells (HUVECs) from a viewpoint of angiogenesis. Sph-1-P facilitated HUVEC spreading on the basement membrane component Matrigel, at concentrations ranging from 10 to 250 nM. This stimulatory response induced by Sph-1-P was blocked by pertussis toxin and C3 transferase (from Clostridium botulinum), which inactivate G(i)-type heterotrimeric G protein and Rho, respectively. Furthermore, Sph-1-P, in the modified Boyden's chamber assay, stimulated HUVEC migration in a concentration-dependent manner, up to 250 nM. Checkerboard analysis revealed that Sph-1-P markedly induces directional migration (chemotaxis), but a random motility (chemokinesis) was also enhanced. The stimulatory effect of Sph-1-P on HUVEC migration was much stronger than that of other bioactive lipids, and again inhibited by pertussis toxin and by C3 transferase. Our present results that Sph-1-P induces endothelial spreading and migration through G(i)-coupled cell surface receptor(s) and Rho are consistent with a recent report on the role of this platelet-derived sphingolipid as a novel regulator of angiogenesis.
Okamoto
H
H
First Department of Surgery, Yamanashi Medical University, Nakakoma, 409-3898, Yamanashi, Japan.
Yatomi
Y
Y
Ohmori
T
T
Satoh
K
K
Matsumoto
Y
Y
Ozaki
Y
Y
D036002:ADP Ribose Transferases / Q000494:pharmacology; D001095:Arachidonic Acids / Q000494:pharmacology; D001792:Blood Platelets / Q000378:metabolism*; D001905:Botulinum Toxins; D002465:Cell Movement / Q000187:drug effects; D048429:Cell Size / Q000187:drug effects; D002478:Cells, Cultured / Q000187:drug effects; D002518:Ceramides / Q000494:pharmacology; D002633:Chemotaxis / Q000187:drug effects; D003094:Collagen; D004338:Drug Combinations; D063388:Endocannabinoids; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D019206:GTP-Binding Protein alpha Subunits, Gi-Go / Q000037:antagonists & inhibitors / Q000502:physiology*; D006801:Humans; D007797:Laminin; D008246:Lysophospholipids / Q000494:pharmacology; D018919:Neovascularization, Physiologic / Q000502:physiology*; D037342:Pertussis Toxin; D015539:Platelet Activation; D053284:Polyunsaturated Alkamides; D011509:Proteoglycans; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000494:pharmacology; D013268:Stimulation, Chemical; D014471:Umbilical Veins; D010566:Virulence Factors, Bordetella / Q000494:pharmacology; D020741:rho GTP-Binding Proteins / Q000037:antagonists & inhibitors / Q000502:physiology*
D001095:Arachidonic Acids; D002518:Ceramides; D004338:Drug Combinations; D063388:Endocannabinoids; D004791:Enzyme Inhibitors; D007797:Laminin; D008246:Lysophospholipids; D053284:Polyunsaturated Alkamides; D011509:Proteoglycans; D010566:Virulence Factors, Bordetella; C059485:matrigel; C060506:sphingosine 1-phosphate; D003094:Collagen; D036002:ADP Ribose Transferases; C063007:exoenzyme C3, Clostridium botulinum; D037342:Pertussis Toxin; D001905:Botulinum Toxins; D019206:GTP-Binding Protein alpha Subunits, Gi-Go; D020741:rho GTP-Binding Proteins; C061800:N,N-dimethylsphingosine; D013110:Sphingosine; C078814:anandamide