LOSS OF PROTEIN-KINASE C-DELTA FROM HUMAN HACAT KERATINOCYTES UPON RAS TRANSFECTION IS MEDIATED BY TGF-ALPHA

The spontaneously immortalized human skin keratinocytes HaCaT contain protein kinase C (PKC)alpha, -delta, -epsilon, and -zeta, All PKC isoenzymes except PKC zeta are down-regulated by TPA as well as by bryostatin, However, with PKC delta, bryostatin but not TPA was found to be much less effective at high concentrations than at low ones. PKC zeta expression at the protein and mRNA level is significantly suppressed in HaCaT cells I-7 and II-4, which are transfected with mutated c-Ha-ras. The expression of the other isoenzymes remains essentially unchanged in the ras-transfected cells compared to normal ones. PKC delta is lost when growing HaCaT cells in a medium obtained from the cultivation of ras-transfected cells (''ras-conditioned'' medium). The factor 0secreted into the medium by the ras-transfected cells that is responsible for this effect appears to be TGF alpha, since the action of ras-conditioned medium on PKC delta expression can be overcome by the addition of an anti-TGF alpha antibody. Moreover, treatment of HaCaT cells with TGF alpha suppresses selectively the expression of the PKC isoenzyme delta. (C) 1995 Academic Press, Inc.