Ras/Raf1-dependent signal in sphingosine-1-phosphate-induced tube formation in human coronary artery endothelial cells.
Since we recently reported that high density lipoprotein, which contains the bioactive lipid sphingosine-1-phosphate (S1P) [Arterioscler. Thromb. Vasc. Biol. 23 (2003) 802], induced human coronary artery endothelial cell (HCEC) tube formation mediated by a Ras/Raf/ERK (extracellular signal-activated kinase) pathway, we thought that it would be very important to evaluate whether the signal in S1P-induced tube formation is Ras-dependent or -independent. In an in vitro model of HCEC tube formation on a matrix gel, S1P-induced tube formation. ERK1/2 inhibitor (PD98059) and pertussis toxin (PTX) suppressed S1P-induced tube formation. S1P activated phospho(p)-ERK1/2, while dominant-negative RasN17 blocked S1P-induced p-ERK1/2. Moreover, RasN17 inhibited S1P-induced tube formation. S1P activated Ras/Raf1 by Ras pull-down assay and this effect was inhibited by PTX. These results demonstrate that Ras/Raf1-dependent ERK activation mediated by PTX-sensitive G protein-coupled receptors may be a potent signal in S1P-induced HCEC tube formation.
Miura
Shin-ichiro
S
Department of Cardiology, Fukuoka University School of Medicine, 7-45-1 Nanakuma, Jonan-Ku, Fukuoka 814-0180, Japan. miuras@cis.fukuoka-u.ac.jp
Tanigawa
Hiroyuki
H
Matsuo
Yoshino
Y
Fujino
Masahiro
M
Kawamura
Akira
A
Saku
Keijiro
K
D002478:Cells, Cultured; D003094:Collagen / Q000494:pharmacology; D003331:Coronary Vessels / Q000166:cytology*; D004305:Dose-Response Relationship, Drug; D004338:Drug Combinations; D004730:Endothelium, Vascular / Q000378:metabolism*; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005419:Flavonoids / Q000494:pharmacology; D005799:Genes, Dominant; D006801:Humans; D015151:Immunoblotting; D007797:Laminin / Q000494:pharmacology; D008246:Lysophospholipids; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors; D037342:Pertussis Toxin / Q000494:pharmacology; D011509:Proteoglycans / Q000494:pharmacology; D019908:Proto-Oncogene Proteins c-raf / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015398:Signal Transduction; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000378:metabolism*; D013997:Time Factors; D014162:Transfection; D018631:ras Proteins / Q000378:metabolism*
D004338:Drug Combinations; D004791:Enzyme Inhibitors; D005419:Flavonoids; D007797:Laminin; D008246:Lysophospholipids; D011509:Proteoglycans; D011993:Recombinant Fusion Proteins; C059485:matrigel; C060506:sphingosine 1-phosphate; D003094:Collagen; D037342:Pertussis Toxin; D019908:Proto-Oncogene Proteins c-raf; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases; D018631:ras Proteins; D013110:Sphingosine; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one