Rapid screening method for inhibitors of protein kinase C.
Specific inhibitors of protein kinase C (PKC) were screened for with a unique detection system, named bleb forming assay. When K562, a human chronic myeloid leukemia cell, was treated with phorbol 12,13-dibutylate (PDBu) or teleocidin which are activators of PKC, many blebs appeared on the cell surface of K562 within 10 minutes. This appearance of blebs is inhibited by staurosporine and H7 which are known to be PKC inhibitors. Teleocidin and PDBu did not induce bleb formation of HL60, a human acute promyelocytic leukemia cell, and the mouse Friend leukemia cell, even though their morphology was changed 24 hours after treatment with teleocidin or PDBu. Many inducers of terminal differentiation of K562 have the same effect on HL60 and Friend cells. However, the bleb inducing activity of PKC activators seems to be specific for K562. The bleb forming assay satisfied the criteria (simplicity and specificity) required for preliminary screening of activators or inhibitors of PKC. Teleocidins A and B, and tautomycin (a new antibiotic isolated in our laboratory) were identified as activators of PKC, and also staurosporine and isoflavones (daidzein and genistein) as inhibitors.
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D000192:Actinomycetales / Q000032:analysis; D000470:Alkaloids / Q000494:pharmacology; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D007529:Isoflavones / Q000494:pharmacology; D007546:Isoquinolines / Q000494:pharmacology; D007951:Leukemia, Myeloid / Q000473:pathology; D008235:Lyngbya Toxins / Q000494:pharmacology; D015240:Phorbol 12,13-Dibutyrate; D010703:Phorbol Esters / Q000494:pharmacology; D010879:Piperazines / Q000494:pharmacology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors*; D019311:Staurosporine; D014407:Tumor Cells, Cultured
D000470:Alkaloids; D000900:Anti-Bacterial Agents; D007529:Isoflavones; D007546:Isoquinolines; D008235:Lyngbya Toxins; D010703:Phorbol Esters; D010879:Piperazines; C022176:teleocidins; D015240:Phorbol 12,13-Dibutyrate; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D011493:Protein Kinase C; D019311:Staurosporine