Tumor necrosis factor (TNF) caused an inhibition of 125I-labeled epidermal growth factor [( 125I]EGF) binding to its receptors of human amniotic (WISH) cells at 5 min after addition of TNF, which reached a maximal level (60-70% reduction) after 15-30 min and declined thereafter. TNF also induced a translocation of protein kinase C activity from the cytosol to the membrane, which peaked at 45-60 min after addition of TNF and almost returned to basal level at 120 min. Furthermore, prolonged incubation of WISH cells with 12-O-tetradecanoylphorbol 13 acetate (TPA) diminished the TPA effect on the inhibition of EGF binding to the cells due to the desensitization of protein kinase C; however, TNF still reduced the EGF binding to the cells pretreated with TPA for a long time. These results indicate that although TNF causes the translocation of protein kinase C to the membrane, activation of protein kinase C is not required for TNF to induce a decrease in EGF binding to the cells.