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  • Title: Differential down-regulation of protein kinase C subspecies in normal human melanocytes: possible involvement of the zeta subspecies in growth regulation.
    Author: Oka M, Ogita K, Ando H, Kikkawa U, Ichihashi M.
    Journal: J Invest Dermatol; 1995 Oct; 105(4):567-71. PubMed ID: 7561160.
    Abstract:
    Normal human melanocytes are often grown in vitro in the continuous presence of 12-O-tetradecanoylphorbol-13-acetate (TPA) for growth in vitro. The expression of protein kinase C (PKC) subspecies, which are the major cellular receptors for phorbol esters, was examined in melanocytes after long-term treatment with TPA to investigate the role of PKC subspecies in TPA-dependent cell growth. The PKC enzyme activity detected in quiescent melanocytes was almost completely depleted in cells after incubation with 85 nM TPA for 48 h. Immunoblot analysis indicated that, among the PKC subspecies alpha, beta, delta, epsilon, and zeta expressed in quiescent cells, alpha-, beta-, delta-, and epsilon-PKC were significantly down-regulated, whereas zeta-PKC remained at detectable levels in TPA-treated cells. TPA did not significantly affect the expression or subcellular distribution of zeta-PKC in melanocytes. Immunoprecipitation assay revealed that the enzyme activity of zeta-PKC was increased in both the cytosol and particulate cell fractions, but the increase was much greater in the latter. The activation of zeta-PKC lasted for 24 to 48 h after the addition of TPA; thereafter, zeta-PKC activity returned to basal levels. DNA synthesis was shown to change concomitantly with the activation of zeta-PKC in TPA-treated cells. These results indicate that TPA induces not only the down-regulation of alpha-, beta-, delta-, and epsilon-PKC, but also long-term activation of zeta-PKC in melanocytes, and that activation of zeta-PKC parallels the growth of normal human melanocytes.
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