Comparison of ability of protein kinase C inhibitors to arrest cell growth and to alter cellular protein kinase C localisation.
Inhibitors of protein kinase C (PKC) such as the staurosporine analogues UCN-01 and CGP 41251 possess antineoplastic properties, but the mechanism of their cytostatic action is not understood. We tested the hypothesis that the ability of these compounds to arrest growth is intrinsically linked with their propensity to inhibit PKC. Compounds with varying degrees of potency and specificity for PKC were investigated in A549 and MCF-7 carcinoma cells. When the log values of drug concentration which arrested cell growth by 50% (IC50) were plotted against the logs of the IC50 values for inhibition of cytosolic PKC activity, two groups of compound could be distinguished. The group which comprised the more potent inhibitors of enzyme activity (calphostin C, staurosporine and its analogues UCN-01, RO 31-8220, CGP 41251) were the stronger growth inhibitors, whereas the weaker enzyme inhibitors (trimethylsphingosine, miltefosine, NPC-15437, H-7, H-7I) affected proliferation less potently. GF 109203X was exceptional in that it inhibited PKC with an IC50 in the 10(-8) M range, yet was only weakly cytostatic. To substantiate the role of PKC in the growth inhibition caused by these agents, cells were depleted of PKC by incubation with bryostatin 1 (1 microM). The susceptibility of these enzyme-depleted cells towards growth arrest induced by staurosporine, RO 31-8220, UCN-01 or H-7 was studied. The drug concentrations which inhibited incorporation of [3H]thymidine into PKC-depleted A549 cells by 50% were slightly, but not significantly, lower than significantly, lower than those observed in control cells. These results suggest that PKC is unlikely to play a direct role in the arrest of the growth of A549 and MCF-7 cells mediated by these agents. Staurosporine is not only a strong inhibitor of PKC but also mimics activators of this enzyme in that it elicits the cellular redistribution of certain PKC isoenzymes. The ability of kinase inhibitors other than staurosporine to exert a similar effect was investigated. Calphostin C, H-7, H-7I, miltefosine, staurosporine, UCN-01, RO 31-8220, CGP 41251 or GF 109203X were incubated for 30 min with A549 cells in the absence or presence of the PKC activator 12-O-tetradecanoyl phorbol-13-acetate. The subcellular distribution of PKC-alpha-, -epsilon and -zeta was measured by Western blot analysis. None of the agents affected PKC-alpha or -zeta.(ABSTRACT TRUNCATED AT 400 WORDS)
Courage
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Medical Research Council Toxicology Unit, University of Leicester, UK.
Budworth
J
J
Gescher
A
A
D000470:Alkaloids / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000494:pharmacology*; D001943:Breast Neoplasms; D002455:Cell Division / Q000187:drug effects*; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D006801:Humans; D007527:Isoenzymes / Q000032:analysis / Q000378:metabolism; D008175:Lung Neoplasms; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*; D019311:Staurosporine / Q000031:analogs & derivatives; D013329:Structure-Activity Relationship; D014407:Tumor Cells, Cultured
D000470:Alkaloids; D000970:Antineoplastic Agents; D007527:Isoenzymes; C054852:7-hydroxystaurosporine; D011493:Protein Kinase C; D019311:Staurosporine; C059539:midostaurin