Although keratinocytes are a major target of phorbol ester actions, the activity and the expression of the eight cloned protein kinase C (PKC) isoenzymes have not been studied in detail in human epidermis. Starting from normal human keratinocytes, we reconstituted in culture a multilayered epithelial tissue which presents many hystological, biochemical, and molecular features of the authentic epidermis and we used it as a model to investigate the PKC activity and mRNA levels. We found that i) PKC activity is higher in differentiated than in non-differentiated cells; ii) the mRNA levels of PKC delta and -eta/L, while are differently affected by spontaneous keratinocyte differentiation, are down-regulated during phorbol esters-induced cell differentiation. Our findings could represent a basis to investigate the involvement of PKC isoforms in the keratinocyte differentiation process.