Research Article| Volume 4, ISSUE 1, P18-25, July 1992

Anti-proliferative effects of protein kinase C inhibitors in human keratinocytes

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      Various lines of evidence indicate that protein kinase C, a key enzyme in transmembraneous signal transduction, is involved in the regulation of keratinocyte proliferation. In the present study we have investigated the effects of various structurally unrelated protein kinase C inhibitors on the proliferation of HaCaT cells, a non-tumorigenic human keratinocyte cell line. All protein kinase C inhibitors dose-dependently inhibited cell proliferation as assessed by the incorporation of radioactively labelled thymidine and amino acids as well as the increase in total protein content in keratinocytes. The potencies of the drugs to inhibit cell proliferation were strongly correlated to their inhibitory potency on purified protein kinase C, displaying a correlation coefficient of 0.97. Methotrexate, an anti-proliferative drug, was found not to inhibit protein kinase C. Therefore, our data provide evidence that protein kinase C is crucially involved in the regulation of keratinocyte proliferation but is not the only target of anti-proliferative drug action.



      CaM (calmodulin), CP 46,665-1 (4-aminomethyl-1-[2,3-(di-n-decyloxy)n-proponyl]-4-phenylpiperidine dihydrochloride), DNA (deoxyribonucleic acid), PMA (phorbol-12-myristate-13-acetate), PKC (protein kinase C), PS (phosphatidyl-l-serine), W-7 (N-(6-aminohexyl)-5-chloro-1-naphthalene sulfonamide)
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        • Fisher GJ
        • Talwar HS
        • Tavakkol A
        • Esmann J
        • Baldassare JJ
        • Elder JT
        • Griffiths CEM
        • Baadsgaard O
        • Cooper KD
        • Vorrhees JJ
        Phosphoinositide-mediated signal transduction in normal and psoriatic epidermis.
        J Invest Dermatol. 1990; 95: 15S-17S
        • Berridge MJ
        • Brown KD
        • Irvine RF
        • Heslop JP
        Phosphoinositides and cell proliferation.
        J Cell Sci. 1985; : 187-198
        • Nishizuka Y
        The role of protein kinase C in cell surface signal transduction and tumour promotion.
        Nature. 1984; 308: 693-698
        • Marks F
        Prostaglandins, cyclic nucleotides and the effect of phorbol ester tumor promotors in mouse skin in vivo.
        Carcinogenesis. 1983; 4: 1465-1470
        • Hawley-Nelson P
        • Stanley JR
        • Schmidt J
        • Gullino M
        • Yuspa SH
        The tumor promotor, 12-O-tetradecanoyl-phorbol-13-acetate, accelerates keratinocyte differentiation and stimulates growth of an unidentified cell type in cultured human epidermis.
        Exp Cell Res. 1982; 137: 155-167
        • Snoek GT
        • Boonstra J
        • Ponec M
        • de Laat SW
        Phorbol ester binding and protein kinase C activity in normal and transformed human keratinocytes.
        Exp Cell Res. 1987; 172: 146-157
        • Roenigk HH
        • Maibach HI
        in: Roenigk HH Maibach HI Psoriasis. Marcel Dekker, New York1991: 563-575
        • Tamaoki T
        • Nomoto H
        • Takahashi I
        • Kato Y
        • Morimoto M
        • Tomita F
        Staurosporine, a potent inhibitor of Math Eq dependent protein kinase.
        Biochem Biophys Res Commun. 1986; 135: 397-402
        • Mazzei GJ
        • Katoh N
        • Kuo JF
        Polymyxin B is a more selective inhibitor for phospholipid-sensitive Ca2+-dependent protein kinase than for calmodulin-sensitive Ca2+-dependent protein kinase.
        Biochem Biophys Res Commun. 1982; 109: 1129-1133
        • Shoji M
        • Vogler WR
        • Kuo JF
        Inhibition of Math Eq protein kinase and phosphorylation of leukemic cell proteins by CP-46,665-1, a novel antineoplastic lipoidal amine.
        Biochem Biophys Res Commun. 1985; 127: 590-595
        • O'Brian CA
        • Ward NE
        Binding of protein kinase C to N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide through its ATP binding site.
        Biochem Pharmacol. 1989; 38: 1737-1742
        • Mori T
        • Takai Y
        • Minakuchi R
        • Yu B
        • Nishizuka Y
        Inhibitory action of chlorpromazine, dibucaine, and other phospholipid-interacting drugs on calcium-activated, phospholipid-dependent protein kinase.
        J Biol Chem. 1980; 255: 8378-8380
        • Wise BC
        • Kuo JF
        Modes of inhibition by acylcarnitides, adriamycin and trifluoperazine of cardiac phospholipid-sensitive calcium-dependent protein kinase.
        Biochem Pharmacol. 1983; 32: 1259-1265
        • Boukamp P
        • Petrussevska RT
        • Breitkreutz D
        • Hornung J
        • Markham A
        • Fusenig NE
        Normal keratinization in a spontaneously immortalized aneuploid human keratinocyte cell line.
        J Cell Biol. 1988; 106: 761-771
        • Hegemann L
        • Fruchtmann R
        • Bonnekoh B
        • Schmidt BH
        • Traber J
        • Mahrle G
        • Müller-Peddinghaus R
        • van Rooijen LAA
        Effects of tiflucarbine as a dual protein kinase C/calmodulin antagonist on proliferation of human keratinocytes and release of reactive oxygen species from human leukocytes.
        Arch Dermatol Res. 1991; 283: 456-460
        • Bradford M
        A rapid and sensitive method for the quantification of microgram quantities of protein using the principle of protein-dye binding.
        Anal Biochem. 1976; 72: 248-254
        • Kikkawa U
        • Minakuchi R
        • Takai Y
        • Nishizuka Y
        Calcium-activated, phospholipid-dependent protein kinase (protein kinase C) from rat brain.
        Methods Enzymol. 1983; 99: 288-298
        • Hegemann L
        • Wevers A
        • Bonnekoh B
        • Mahrle G
        Changes of epidermal cell morphology and keratin expression induced by inhibitors of protein kinase C.
        J Dermatol Sci. 1992; 3: 103-110
        • Reymann KG
        • Frey U
        • Jork R
        • Matthies H
        Polymyxin B, an inhibitor of protein kinase C, prevents the maintenance of synaptic long-term potentiation in hippocampal CA1 neurons.
        Brain Res. 1988; 440: 305-314
        • Balinga BS
        • Sindel LJ
        • Jenkins LD
        • Sachen JD
        Effects of polymyxin B on T-lymphocyte protein synthesis.
        Biochem Biophys Res Commun. 1986; 135: 649-654
        • Rüegg UT
        • Burgess GM
        Staurosporine, K-252 and UCN-01: potent but nonspecific inhibitors of protein kinases.
        Trends Pharmacol Sci. 1989; 10: 218-220
        • Hegemann L
        • van Rooijen LAA
        • Traber J
        • Schmidt BH
        Polymyxin B is a selective and potent antagonist of calmodulin.
        Eur J Pharmacol. 1991; 207: 17-22
        • Hidaka H
        • Asano M
        • Tanaka T
        Activity-structure relationship of calmodulin antagonists — naphthalenesulfonamide derivatives.
        Mol Pharmacol. 1981; 20: 571-578
        • Massom L
        • Lee H
        • Jarrett HW
        Trifluoperazine binding to porcine brain calmodulin and skeletal muscle troponin C.
        Biochemistry. 1990; 29: 671-681
        • Yanagihara N
        • Tachikawa E
        • Izumi F
        • Yasugawa S
        • Yamamoto H
        • Miyamoto E
        Staurosporine: an effective inhibitor for Ca2+/calmodulin-dependent protein kinase II.
        J Neurochem. 1991; 56: 294-298
        • Nishizuka Y
        Studies and perspectives of protein kinase C.
        Science. 1986; 233: 305-312
        • Hegemann L
        • Mahrle G
        Biochemical pharmacology of protein kinase C and its relevance for dermatology.
        in: Mukhtar H Pharmacology of the Skin. CRC Press, Boca Raton1991: 357-368
        • Ochs DL
        • Reed PW
        Inhibition of neutrophil oxidative burst and degranulation by phenothiazines.
        Biochem Biophys Res Commun. 1981; 102: 958-962
        • Fujita I
        • Takeshige K
        • Minakami S
        Inhibition of neutrophil superoxide formation by 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H-7), an inhibitor of protein kinase C.
        Biochem Pharmacol. 1986; 35: 4555-4562
      1. Hegemann L, Fruchtmann R, van Rooijen LAA, Müller-Peddinghaus R, Mahrle G: The antipsoriatic drug, anthralin inhibits protein kinase C — implications for its mechanism of action. Arch Dermatol Res (in press).