Potentiation of insulin-induced phosphatidylinositol-3 kinase activity by phorbol ester is mediated by protein kinase C epsilon.

Our previous results have demonstrated that phorbol 12-myristate 13-acetate (TPA) and insulin synergistically stimulate the activity of phosphatidylinositol-3 kinase (PI-3 kinase) and PI-3 kinase plays an important role in both of TPA-induced AP-1 activation and cell transformation in tumour promotion sensitive (P+) JB6 cells. In the present study, we investigated the role of PKC and its isozymes in the synergistic induction of PI-3 kinase by TPA and insulin. Bisindolylmaleimide inhibits TPA- and TPA+ insulin-induced PI-3 kinase activity. Pretreatment of cells for 24 h with TPA has significant inhibitory effects on TPA-induced PI-3 kinase activity and abolishes the synergistic effect of TPA and insulin-stimulated PI-3 kinase activity. Furthermore, overexpression of a dominant negative PKC epsilon, but not dominant negative PKC alpha, blocks the synergistic effect of TPA and insulin-induced PI-3 kinase activity. These results indicate that the potentiation effect of TPA on insulin-induced PI-3 kinase activity is specific through PKC epsilon in JB6 cells.