Long-wave ultraviolet radiation induces protein kinase C in normal human keratinocytes.
Skin tumor promotion by phorbol ester is believed to be mediated by the phospholipid-dependent ser/thr kinase, protein kinase C (PKC). Long-wave ultraviolet radiation (320-400 nm, UVA), which has also been shown to promote skin tumors, induces elevated levels of PKC in murine fibroblasts, suggesting that UVA may promote the development of basal and squamous cell skin cancers by a mechanism involving PKC. To examine UVA effects on PKC in a model relevant to skin, we maintained normal human epidermal keratinocytes (NHEK) in serum-free medium and exposed the cultured cells to various doses of UVA or to the phorbol ester, 12-O-tetradecanoylphorbol-13-acetate (TPA). Fifty minutes after exposure to UVA (5-20 J/cm2), PKC activity was elevated up to three-fold in NHEK cytosolic fractions, and membrane-associated PKC activity was elevated up to two-fold by UVA. The TPA treatment induced a 10-fold increase in membrane-associate PKC activity only. Immunoblot analysis suggested that a UVA-induced increase in PKC protein occurred. Both UVA and TPA reduced the cell number by 50-75% in the first 24-48 h; however, irradiated cultures began to recover at 72 h post-UVA due to an increased proliferative rate beginning after 48 h. Treatment with TPA induced a high level of differentiation as measured by cornified envelope formation. Ultraviolet A irradiation exposure was not followed by increased differentiation. These findings suggest that acute UVA exposure elevates PKC activity in human keratinocytes and may act through PKC to promote actinic skin cancer. The molecular mechanism is like to differ from that of the phorbol esters, however.
Matsui
M S
MS
Department of Dermatology, Columbia University, New York, NY 10032.
Wang
N
N
MacFarlane
D
D
DeLeo
V A
VA
D000595:Amino Acid Sequence; D002454:Cell Differentiation / Q000528:radiation effects; D002455:Cell Division / Q000528:radiation effects; D002478:Cells, Cultured; D004790:Enzyme Induction / Q000528:radiation effects; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000201:enzymology / Q000528:radiation effects*; D008969:Molecular Sequence Data; D011493:Protein Kinase C / Q000096:biosynthesis / Q000528:radiation effects*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013936:Thymidine / Q000378:metabolism; D014466:Ultraviolet Rays
D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate; D013936:Thymidine