BFGF and PDGF-BB have a synergistic effect on the proliferation, migration and VEGF release of endothelial progenitor cells

Abstract

We have investigated the synergistic effects of bFGF (basic fibroblast growth factor) and PDGF-BB (platelet-derived growth factor-BB) on the proliferation, migration and VEGF (vascular endothelial growth factor) release of EPCs (endothelial progenitor cells). The proliferation of EPCs was assayed by MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium]. EPCs migration was detected using the Transwell system. Real-time PCR was used to assess the transcription of PDGFRβ mRNA. PLC-γ (phospholipase C gamma) expression and VEGF release were analysed by Western blot and ELISA. bFGF and PDGF-BB could, respectively, or synergistically, promote the proliferation and migration of EPCs, and these effects of bFGF and PDGF-BB were implemented by enhancing PDGFRβ mRNA, PLC-γ and VEGF expression, while inhibitor of PDGF receptor kinase (AG1296) and the selective PLC inhibitor (U73122) could block these effects of bFGF and PDGF-BB. In the meantime, we proved that the amplification by bFGF and PDGF-BB-stimulated PDGFRβ mRNA, PLC-γ and VEGF expression was abrogated by anti-bFGF antibody, AG1296 and U73122. These results strongly suggest that the proliferation and migration of EPCs may depend on bFGF and/or PDGF-BB by PDGFRβ/PLC-γ signalling pathway, and bFGF and/or PDGF-BB stimulate VEGF release at a point downstream from PDGFRβ/PLC-γ in EPCs.