P70 S6 kinase negatively regulates fibroblast growth factor 2-stimulated interleukin-6 synthesis in osteoblasts: function at a point downstream from protein kinase C

Abstract

We have previously reported that protein kinase C negatively regulates basic fibroblast growth factor (FGF-2)-stimulated synthesis of interleukin-6 (IL-6), a potent bone resorptive agent, in osteoblast-like MC3T3-E1 cells. To further clarify the mechanism underlying the synthesis of IL-6 in osteoblasts, we investigated whether p70 S6 kinase is involved in the FGF-2-stimulated IL-6 synthesis in these cells. Rapamycin, an inhibitor of p70 S6 kinase, significantly enhanced the FGF-2-stimulated IL-6 synthesis in a dose-dependent manner. Downregulation of p70 S6 kinase by siRNA markedly amplified the FGF-2-stimulated IL-6 synthesis. 12-O-Tetradecanoylphorbol-13-acetate (TPA), a direct activator of protein kinase C, induced the phosphorylation of p70 S6 kinase. Go6976 and bisindolylmaleimide I, inhibitors of protein kinase C, suppressed the TPA-stimulated phosphorylation of p70 S6 kinase. Additionally, protein kinase C inhibitors markedly reduced the phosphorylation of p70 S6 kinase induced by FGF-2. These results strongly suggest that p70 S6 kinase functions at a point downstream of protein kinase C and limits the FGF-2-stimulated IL-6 synthesis in osteoblasts.