Multiple pathways lead to activation of the survival mechanism in quiescent BALB/c-3T3 cells.

The survival of density-arrested quiescent murine BALB/c-3T3 cells in serum-free Dulbecco's medium requires the presence of cell growth factors or second messenger agonists. The protein synthesis inhibitor anisomycin blocks the survival-mediating action of the basic fibroblast growth factor (bFGF) and of 12-0-tetradecanoylphorbol 13-acetate (TPA), but has little or no effect on the protective action of platelet-derived growth factor or 8-bromoadenosine 3':5'-cyclic monophosphate (Br-cAMP). The effects of anisomycin are concentration dependent in the range from 2.5 to 25 muM and show that the survival-enhancing abilities of bFGF and TPA critically require protein synthesis, whereas those of platelet-derived growth factor and Br-cAMP do not. The survival-mediating action of bFGF and TPA can also be blocked with the RNA synthesis inhibitors actinomycin D and 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB), whereas the action of platelet-derived growth factor and Br-cAMP is largely resistant. Results on the time course of action of DRB, a selective inhibitor of the synthesis of mRNA precursor molecules, suggest that the RNA required for the survival-enhancing action of bFGF and TPA is present in cells at the time of serum withdrawal and addition of the survival factor and has a half-life greater than 3 h. The new evidence provides further support for the hypothesis that protection of serum-deprived, density-arrested BALB/c-3T3 fibroblasts against death can be achieved either via pathways that entail the synthesis of protein and RNA (e.g., via diacylglycerol-protein kinase C) or via pathways that do not involve de novo biosynthesis (e.g., via cAMP-protein kinase A).