Effect of protein kinase inhibitors on IL-8/NAP-1 release from human umbilical vein endothelial cells.
Several protein kinase inhibitors (PKIs) were investigated for their effects on IL-1 beta, TNF alpha and PMA-induced IL-8 production from human umbilical vein endothelial cells (HUVEC). IL-1 beta (ED50 0.07 ng/ml), TNF alpha (ED50 100 ng/ml) and PMA (ED50 20 ng/ml) induced IL-8 production that could be detected as early as 2 h following stimulation. Staurosporine, a potent but non-specific inhibitor of protein kinases, inhibited PMA-induced (IC50 2 nM) but not IL-1 beta or TNF alpha (IC50 > 200 nM) induced IL-8 production. Neither the cAMP-dependent PKI, KT5720, nor the tyrosine PKIs, genistein, tyrphostin (1-100 microM) or lavendustin A (0.0001-1 microM), inhibited IL-8 production elicited by IL-1 beta. However, the macrolide protein kinase inhibitor geldanamycin (IC50 = 30 nM), but not the closely related analog herbimycin A (5-500 nM), inhibited IL-8 production by 60%. Northern blot analysis of IL-8 mRNA revealed that staurosporine suppressed mRNA increase following stimulation by PMA but not by IL-1. It is proposed that a novel protein kinase susceptible to geldanamycin inhibition may be involved in IL-1-mediated signal transduction.
White
J R
JR
Department of Cellular Biochemistry L101, SmithKline Beecham Pharmaceuticals, King of Prussia, PA 19406.
Lee
J C
JC
D000470:Alkaloids / Q000494:pharmacology; D016227:Benzoquinones; D002227:Carbazoles; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D007375:Interleukin-1 / Q000494:pharmacology*; D016209:Interleukin-8 / Q000378:metabolism*; D047029:Lactams, Macrocyclic; D047428:Protein Kinase Inhibitors; D011758:Pyrroles / Q000494:pharmacology; D011809:Quinones / Q000494:pharmacology; D011994:Recombinant Proteins / Q000494:pharmacology; D017828:Rifabutin / Q000031:analogs & derivatives; D019311:Staurosporine; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*; D014471:Umbilical Veins / Q000187:drug effects / Q000378:metabolism
D000470:Alkaloids; D016227:Benzoquinones; D002227:Carbazoles; D007211:Indoles; D007375:Interleukin-1; D016209:Interleukin-8; D047029:Lactams, Macrocyclic; D047428:Protein Kinase Inhibitors; D011758:Pyrroles; D011809:Quinones; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; D017828:Rifabutin; C057416:KT 5720; C020754:herbimycin; D019311:Staurosporine; D013755:Tetradecanoylphorbol Acetate; C001277:geldanamycin