We have examined the effect of protein kinase C (PKC) on the expression of the E-selectin and intercellular adhesion molecule-1 (ICAM-1) mRNAs in human umbilical vein endothelial cells. The lower classic PKC activity on pretreatment with phorbol ester (phorbol 12-myristate 13-acetate (PMA)) for 24 h markedly decreased IL-1beta-induced E-selectin mRNA expression in the presence of fetal calf serum and basic fibroblast growth factor, although the induction of ICAM-1 mRNA expression was only influenced a little by the PKC down-regulation. On the other hand, tumor necrosis factor-alpha (TNFalpha)-induced gene expression of these adhesion molecules was unaffected by such PKC modulation. The intracellular signals generated by interleukin (IL)-1beta and TNFalpha themselves are not mediated through classic PKC activation, because the response to neither stimulant was inhibited by the PKC down-regulation in the absence of fetal calf serum and basic fibroblast growth factor. Simultaneous treatment with IL-1beta and PMA synergistically induced E-selectin gene expression but not when TNFalpha was substituted for IL-1beta. ICAM-1 mRNA expression was only additively induced on the cotreatment. The synergistic effect on E-selectin mRNA induction was independent of de novo protein synthesis and mediated by elevated transcriptional activity. Promoter analysis of E-selectin indicated that the NF-ELAM1/activating transcription factor element is critical for the synergistic effect of the cotreatment with IL-1beta and PMA.