Mechanisms of fibroblast growth factor-2 modulation of vascular endothelial growth factor expression by osteoblastic cells

Normal bone growth and repair is dependent on angiogenesis. Fibroblast grow th factor-2 (FGF-2), vascular endothelial growth factor (VEGF), and transfo rming growth factor-beta (TGF beta) have all been implicated in the related processes of angiogenesis, growth, development, and repair. The purpose of this study was to investigate the relationships between FGF-2 and both VEG F and TGF beta in nonimmortalized and clonal osteoblastic cells. Northern b lot analysis revealed 6-fold peak increases in VEGF mRNA at 6 h in fetal ra t calvarial cells and MC3T3-E1 osteoblastic cells after stimulation with FG F-S. Actinomycin D inhibited these increases in VEGF mRNA, whereas cyclohex imide did not. The stability of VEGF mRNA was not increased after FGF-2 tre atment. Furthermore, FGF-2 induced dose-dependent increases in VEGF protein levels (P < 0.01). Although in MC3T3-E1 cells, TGF beta 1 stimulates a 6-f old peak increase in VEGF mRNA after 3 h of stimulation, we found that both TGF beta 2 and TGF beta 3 yielded 2- to 8-fold peak increases in VEGF mRNA levels noted after 6 h of stimulation. Similarly, both TGF beta 2 and TCF beta 3 dose dependently increased VEGF protein production. To determine whe ther FGF-2-induced increases in VEGF mRNA may have occurred independently o f TGF beta, we disrupted TGF beta signal transduction (using adenovirus enc oding a truncated form of TGF beta receptor II), which attenuated TGF beta 1 induction of VEGF mRNA, but did not impede FGF-2 induction of VEGF mRNA. In summary, FGF-2-induced VEGF expression by osteoblastic cells is a dose-d ependent event that may be independent of concomitant FGF-2-induced modulat ion of TGF beta activity.