We have recently shown that IFN alpha induces a cytolytic mechanism in human ovarian carcinoma cell lines which is revealed when protein synthesis is subsequently inhibited. In order to determine whether the cytolytic activity induced by IFN alpha was activated through a pathway involving the activation of PKC, the human ovarian carcinoma cell line Caov-3 was exposed to phorbol 12-myristate 13-acetate (PMA). Under conditions where PMA activates PKC, PMA mimicked IFN alpha in its ability to induce a cytolytic mechanism. In contrast, under conditions where PMA depletes PKC, PMA not only did not induce cytolytic activity, but it prevented IFN alpha from inducing cytolytic activity. To further investigate the involvement of PKC in the signaling of cytolytic activity by IFN alpha, the ability of the protein kinase inhibitors, staurosporine, 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine dihydrochloride (H7), N-[2-(methylamino) ethyl]-5-isoquinolinesulfonamide dihydrochloride (H8), and N-(2-guanidinoethyl)-5-isoquinolinesulfonamide dihydrochloride (HA1004) to block the induction of cytolytic activity by IFN alpha was determined. The fact that H7, H8, and staurosporine, but not HA1004, blocked the induction of cytolytic activity by IFN alpha provides additional evidence of the involvement of PKC in this activity. Taken together these results indicate that the cytolytic activity induced by IFN alpha is induced through apathway that involves the activation of PKC.