TNF-alpha-induced cyclooxygenase-2 expression in human lung epithelial cells: involvement of the phospholipase C-gamma 2, protein kinase C-alpha, tyrosine kinase, NF-kappa B-inducing kinase, and I-kappa B kinase 1/2 pathway.
TNF-alpha induced a dose- and time-dependent increase in cyclooxygenase-2 (COX-2) expression and PGE2 formation in human NCI-H292 epithelial cells. Immunofluorescence staining demonstrated that COX-2 was expressed in cytosol and nuclear envelope. Tyrosine kinase inhibitors (genistein or herbimycin) or phosphoinositide-specific phospholipase C inhibitor (U73122) blocked TNF-alpha-induced COX-2 expression. TNF-alpha also stimulated phosphatidylinositol hydrolysis and protein kinase C (PKC) activity, and both were abolished by genistein or U73122. The PKC inhibitor, staurosporine, also inhibited TNF-alpha-induced response. The 12-O-tetradecanoylphorbol 13-acetate (TPA), a PKC activator, also stimulated COX-2 expression, this effect being inhibited by genistein or herbimycin. NF-kappaB DNA-protein binding and COX-2 promoter activity were enhanced by TNF-alpha, and these effects were inhibited by genistein, U73122, staurosporine, or pyrolidine dithiocarbamate. TPA stimulated both NF-kappaB DNA-protein binding and COX-2 promoter activity, these effects being inhibited by genistein, herbimycin, or pyrolidine dithiocarbamate. The TNF-alpha-induced, but not the TPA-induced, COX-2 promoter activity was inhibited by phospholipase C-gamma2 mutants, and the COX-2 promoter activity induced by either agent was attenuated by dominant-negative mutants of PKC-alpha, NF-kappaB-inducing kinase, or I-kappaB (inhibitory protein that dissociates from NF-kappaB) kinase (IKK)1 or 2. IKK activity was stimulated by both TNF-alpha and TPA, and these effects were inhibited by staurosporine or herbimycin. These results suggest that, in NCI-H292 epithelial cells, TNF-alpha might activate phospholipase C-gamma2 via an upstream tyrosine kinase to induce activation of PKC-alpha and protein tyrosine kinase, resulting in the activation of NF-kappaB-inducing kinase and IKK1/2, and NF-kappaB in the COX-2 promoter, then initiation of COX-2 expression and PGE2 release.
Chen
C C
CC
Institute of Pharmacology, College of Medicine, National Taiwan University, Taipei, Taiwan. ccchen@ha.mc.ntu.edu.tw
Sun
Y T
YT
Chen
J J
JJ
Chiu
K T
KT
D016227:Benzoquinones; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D051546:Cyclooxygenase 2; D004789:Enzyme Activation / Q000187:drug effects / Q000235:genetics / Q000276:immunology; D004790:Enzyme Induction / Q000187:drug effects / Q000276:immunology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004847:Epithelial Cells / Q000187:drug effects / Q000201:enzymology* / Q000276:immunology / Q000378:metabolism; D004963:Estrenes / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000276:immunology; D019833:Genistein / Q000494:pharmacology; D006801:Humans; D051550:I-kappa B Kinase; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D047029:Lactams, Macrocyclic; D008168:Lung / Q000187:drug effects / Q000201:enzymology* / Q000276:immunology / Q000378:metabolism; D008565:Membrane Proteins; D016297:Mutagenesis, Site-Directed; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D051966:Phospholipase C gamma; D011401:Promoter Regions, Genetic / Q000276:immunology; D011451:Prostaglandin-Endoperoxide Synthases / Q000096:biosynthesis* / Q000235:genetics; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000502:physiology*; D051571:Protein Kinase C-alpha; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000502:physiology*; D011759:Pyrrolidines / Q000494:pharmacology; D011760:Pyrrolidinones / Q000494:pharmacology; D011809:Quinones / Q000494:pharmacology; D017828:Rifabutin / Q000031:analogs & derivatives; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics / Q000276:immunology*; D019311:Staurosporine / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013859:Thiocarbamates / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors / Q000502:physiology*; D010738:Type C Phospholipases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D000097782:NF-kappaB-Inducing Kinase
D016227:Benzoquinones; D051546:Cyclooxygenase 2; D004791:Enzyme Inhibitors; D004963:Estrenes; D019833:Genistein; D051550:I-kappa B Kinase; D007527:Isoenzymes; D047029:Lactams, Macrocyclic; D008565:Membrane Proteins; D016328:NF-kappa B; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D051966:Phospholipase C gamma; D011451:Prostaglandin-Endoperoxide Synthases; D011493:Protein Kinase C; D051571:Protein Kinase C-alpha; D017346:Protein Serine-Threonine Kinases; D011505:Protein-Tyrosine Kinases; D011759:Pyrrolidines; D011760:Pyrrolidinones; D011809:Quinones; D017828:Rifabutin; D019311:Staurosporine; D013755:Tetradecanoylphorbol Acetate; D013859:Thiocarbamates; D014409:Tumor Necrosis Factor-alpha; D010738:Type C Phospholipases; D000097782:NF-kappaB-Inducing Kinase; C060229:1-(6-((3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione; C020972:pyrrolidine dithiocarbamic acid; C020754:herbimycin; C496540:PTGS2 protein, human; C496559:CHUK protein, human; C496560:IKBKB protein, human; C496565:IKBKE protein, human; C496628:PRKCA protein, human