Human Fetal Skin Fibroblast Migration Stimulated by the Autocrine Growth Factor bFGF Is Mediated by Phospholipase A2 via Arachidonic Acid without the Involvement of Pertussis Toxin-Sensitive G-Protein

Abstract

We reported previously that human fetal skin fibroblast migration into a denuded area was stimulated by an autocrine factor, basic fibroblast growth factor (bFGF). Since the signal transduction pathway of this migration is unknown, we attempted to clarify it by comparing this fibroblast migration with a previously reported bovine endothelial cell migration into a wounded area stimulated by an addition of bFGF, in which the bFGF signal was mediated by phospholipase A2-coupled G-protein and phospholipase A2 (PLA2) via arachidonic acid. Our study demonstrated that pertussis toxin, a specific inhibitor of PLA2-coupled G-protein, did not suppress human fetal skin fibroblast migration, but 2-(p-amylcinnamyl)amino-4-chlorobensoic acid (ONO-RS-082), a PLA2 inhibitor, did. Since ONO-RS-082 is a non-specific PLA2 inhibitor, a cytoplasmic, Ca-dependent PLA2 (cPLA2) inhibitor, AACOCF3, was examined. AACOCF3 suppressed cell migration in certain concentrations. The PLA2 inhibitor-suppressed cell migration was restored by adding arachidonic acid, and cell migration suppressed by anti-bFGF antibodies was restored by adding arachidonic acid. In addition, pertussis toxin did not suppress arachidonic acid release, which shows an action of PLA2, but AACOCF3 did. These results indicate that human fetal skin fibroblast migration stimulated by an autocrine factor, bFGF, was mediated by PLA2 via arachidonic acid without the involvement of PLA2-coupled G-protein.