FGF2-stimulated release of endogenous FGF1 is associated with reduced apoptosis in retinal pigmented epithelial cells

Both inhibition of endogenous fibroblast growth factor (FGF) synthesis on nondividing lens epithelial cells and inhibition of secreted FGF1 in confluent quiescent retinal pigmented epithelial (RPE) cells induce rapid cell apoptosis (Renaud et al., 1996, J. Biol. Chem., 271, 2801-2811). In addition several studies demonstrate that exogenous FGF2 can promote retinal cell survival in vitro and in vivo. To determine the possible relationship between exogenous FGF2, endogenous FGF1, and cell survival, we examined the protective effect of a single dose of exogenous FGF2 on long-term culture of quiescent RPE cells after serum withdrawal. After 4 days of culture, a dramatic and sustained upregulation of FGF1 protein expression occurs specifically in response to exogenous FGF2. After addition of FGF2 (20 ng/ml), RPE cells express fourfold more FGF1 after Day 7 than after Day 1 of culture. This phenomenon is FGF2 dose-dependent. In contrast, neither serum nor FGF2 have an effect on total endogenous FGF2 expression. In addition, in response to exogenous FGF2, FGF1 is secreted in significant amounts into the extracellular medium at a rate comparable to FGF1 accumulation within the cell. Furthermore, in the absence of serum, significant increase in cell death occurs on Day 6 of culture, whereas addition of exogenous FGF2 induces a twofold decrease of RPE cell apoptosis. In the presence of exogenous FGF2, addition of a specific anti-FGF1 neutralizing antibody induces a rapid apoptosis of RPE cell cultures. Thus, we speculate that exogenous FGF2 may indirectly prolong cell survival by increasing synthesis and secretion of endogenous FGF1 and that endogenous FGF1, directly in response to exogenous FGF2, may function as an autocrine trophic factor in RPE cells.