Activation of PKCdelta and p38delta MAPK during okadaic acid dependent keratinocyte apoptosis.
There is substantial interest in identifying agents that differentially activate keratinocyte differentiation versus apoptosis. Okadaic acid (OA) is a tumor promoter in mouse skin that also stimulates apoptosis of murine keratinocytes. OA also enhances human keratinocyte differentiation; however, the impact of OA treatment on apoptosis in these cells has not been examined. We show that OA promotes normal human keratinocyte apoptosis as evidenced by increased accumulation of cells having sub-G1/S DNA content, decreased mitochondrial integrity, increased annexin V binding, increased cytoplasmic cytochrome c level, and increased procaspase 3 and PARP cleavage. Cyclin A, cyclin D1, cdk2, cdk4, p53 and p21 levels are reduced. These changes are associated with release of the PKCdelta catalytic domain and increased phosphorylation of PKCdelta-T(505)-responses consistent with PKCdelta activation. In contrast, phosphorylation of PKCdelta-Y(311) is not increased. The apoptotic response is enhanced in OA treated cells in the presence of p38delta, a PKCdelta target. OA treatment selectively activated p38delta, and OA-dependent apoptosis is not inhibited by treatment with the p38alpha/beta inhibitor, SB203580. These findings are consistent with the idea that the response is mediated by p38delta. Our data indicate that OA is an agent that regulates both keratinocyte differentiation and apoptosis, and that this regulation is mediated via activation of a PKCdelta/p38delta signaling cascade.
Kraft
Catherine A
CA
Department of Physiology and Biophysics, Case Western Reserve University School of Medicine, Cleveland, OH, 44106-4970, USA.
Efimova
Tatiana
T
Eckert
Richard L
RL
D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D018797:Cell Cycle Proteins / Q000187:drug effects / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D004789:Enzyme Activation / Q000502:physiology; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D006801:Humans; D015603:Keratinocytes / Q000187:drug effects / Q000201:enzymology* / Q000473:pathology; D008297:Male; D008928:Mitochondria / Q000187:drug effects / Q000502:physiology; D048292:Mitogen-Activated Protein Kinase 13 / Q000378:metabolism*; D019319:Okadaic Acid / Q000494:pharmacology*; D051745:Protein Kinase C-delta / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology
D018797:Cell Cycle Proteins; D004791:Enzyme Inhibitors; D019319:Okadaic Acid; D048292:Mitogen-Activated Protein Kinase 13; D051745:Protein Kinase C-delta