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Papers In Press, published online ahead of print April 8, 2002
J. Biol. Chem, 10.1074/jbc.M110955200
Submitted on November 15, 2001
Revised on April 4, 2002
Accepted on April 5, 2002

Phosphorylation of STAT-3 in response to basic fibroblast growth factor occurs through a mechanism involving platelet-activating factor, JAK-2 and Src in human umbilical vein endothelial cells: Evidence for a dual kinase mechanism

Dayanand D. Deo, T. William Axelrad, Everett G. Robert, Victor Marcheselli, Nicolas G. Bazan, and Jay D. Hunt

Department of Biochemistry and Molecular Biology, LSU Health Sciences Center, New Orleans, LA 70112

Corresponding Author: jhunt{at}lsuhsc.edu

Platelet-activating factor (PAF1) is a potent proinflamatory phospholipid with multiple pathological and physiological effects. We have shown that bFGF supplementation induces rapid proliferation of human umbilical vein endothelial cells (HUVEC), which is reduced upon removal of bFGF, or by bFGF immunoneutralization. The PAF receptor antagonist LAU-8080 inhibited bFGF-stimulated HUVEC proliferation, indicating the involvement of PAF in the bFGF-mediated signaling of HUVEC. Although FGF receptor (FGFR) phosphorylation was not affected by LAU-8080, the bFGF-mediated prolonged phosphorylation and activation of Erk-1 and –2 were attenuated. Phosphorylation of STAT-3 was observed in the presence of PAF or bFGF, which was attenuated by PAFR antagonists. PAF-induced STAT-3 phosphorylation observed in HUVEC pretreated with either Src inhibitor PP1 or JAK-2 inhibitor AG-490 indicated: (i) immediate (1-minute) phosphorylation of STAT-3 is dependent on Src, (ii) JAK-2 dependent STAT-3 phosphorylation occurs after the delayed (30-minute) PAF exposure, and (iii) prolonged (60-minute) STAT-3 phosphorylation may be either through Src and/or JAK-2. Attenuation of the STAT-3 phosphorylation by the PAFR antagonists indicated signaling through the PAFR. Taken together, these findings suggest the production of PAF is important for bFGF-mediated signaling and that a dual kinase mechanism is involved in the PAF-mediated signal transduction cascade.


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