title: Signal pathways JNK and NF-kappaB, identified by global gene expression profiling, are involved in regulation of TNFalpha-induced mPGES-1 and COX-2 expression in gingival fibroblasts.
abstract: BACKGROUND
  Prostaglandin E2 (PGE2) is involved in several chronic inflammatory diseases including periodontitis, which causes loss of the gingival tissue and alveolar bone supporting the teeth. We have previously shown that tumor necrosis factor alpha (TNFalpha) induces PGE2 synthesis in gingival fibroblasts. In this study we aimed to investigate the global gene expression profile of TNFalpha-stimulated primary human gingival fibroblasts, focusing on signal pathways related to the PGE2-synthesizing enzymes prostaglandin E synthases (PGES), as well as the upstream enzyme cyclooxygenase-2 (COX-2) and PGE2 production.
  
  
  RESULTS
  Microarray and western blot analyses showed that the mRNA and protein expression of the inflammatory induced microsomal prostaglandin E synthase-1 (mPGES-1) was up-regulated by the cytokine TNFalpha, accompanied by enhanced expression of COX-2 and increased production of PGE2. In contrast, the expression of the isoenzymes microsomal prostaglandin E synthase-2 (mPGES-2) and cytosolic prostaglandin E synthase (cPGES) was unaffected by TNFalpha treatment. Using oligonucleotide microarray analysis in a time-course factorial design including time points 1, 3 and 6 h, differentially expressed genes in response to TNFalpha treatment were identified. Enrichment analysis of microarray data indicated two positively regulated signal transduction pathways: c-Jun N-terminal kinase (JNK) and Nuclear Factor-kappaB (NF-kappaB). To evaluate their involvement in the regulation of mPGES-1 and COX-2 expression, we used specific inhibitors as well as phosphorylation analysis. Phosphorylation analysis of JNK (T183/Y185) and NF-kappaB p65 (S536) showed increased phosphorylation in response to TNFalpha treatment, which was decreased by specific inhibitors of JNK (SP600125) and NF-kappaB (Bay 11-7082, Ro 106-9920). Inhibitors of JNK and NF-kappaB also decreased the TNFalpha-stimulated up-regulation of mPGES-1 and COX-2 as well as PGE2 production.
  
  
  CONCLUSIONS
  In the global gene expression profile, the enrichment analysis of microarray data identified the two signal transduction pathways JNK and NF-kappaB as positively regulated by the cytokine TNFalpha. Inhibition of these TNFalpha-activated signal pathways reduced the expression of mPGES-1 and COX-2 as well as their end product PGE2 in gingival fibroblasts. The involvement of the signal pathways JNK and NF-kappaB in the regulation of PGE2 induced by TNFalpha may suggest these two pathways as possible attractive targets in the chronic inflammatory disease periodontitis.
authors.affiliation: Division of Pediatric Dentistry, Department of Dental Medicine, Karolinska Institutet, Huddinge, Sweden. tove.bage@ki.se
authors.forename: Tove
authors.initials: T
authors.lastname: Båge
authors.forename: Johan
authors.initials: J
authors.lastname: Lindberg
authors.forename: Joakim
authors.initials: J
authors.lastname: Lundeberg
authors.forename: Thomas
authors.initials: T
authors.lastname: Modéer
authors.forename: Tülay
authors.initials: T
authors.lastname: Yucel-Lindberg
mesh_terms: D051546:Cyclooxygenase 2 / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism*; D020869:Gene Expression Profiling; D005881:Gingiva / Q000378:metabolism*; D006801:Humans; D019746:Intramolecular Oxidoreductases / Q000378:metabolism*; D052002:NF-kappa B p50 Subunit / Q000378:metabolism*; D000071876:Prostaglandin-E Synthases; D015398:Signal Transduction; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*
chemical_list: D052002:NF-kappa B p50 Subunit; C489548:NFKB1 protein, human; D014409:Tumor Necrosis Factor-alpha; D051546:Cyclooxygenase 2; D019746:Intramolecular Oxidoreductases; C000605862:PTGES protein, human; C000605863:PTGES2 protein, human; D000071876:Prostaglandin-E Synthases
references.citation: J Biol Chem. 2004 May 21;279(21):22057-65
references.pmid: 15023995
references.citation: Exp Eye Res. 2007 Aug;85(2):280-8
references.pmid: 17604021
references.citation: Lab Invest. 2005 Feb;85(2):225-36
references.pmid: 15531909
references.citation: Naunyn Schmiedebergs Arch Pharmacol. 2009 Aug;380(2):153-60
references.pmid: 19415240
references.citation: Nucleic Acids Res. 2007 Jan;35(Database issue):D760-5
references.pmid: 17099226
references.citation: Nucleic Acids Res. 2007 Jan;35(Database issue):D26-31
references.pmid: 17148475
references.citation: Inflammation. 2004 Apr;28(2):89-95
references.pmid: 15379214
references.citation: J Biol Chem. 1997 Aug 22;272(34):21096-103
references.pmid: 9261113
references.citation: J Periodontal Res. 2003 Apr;38(2):198-203
references.pmid: 12608915
references.citation: Biochim Biophys Acta. 1999 Aug 18;1439(3):406-14
references.pmid: 10446427
references.citation: J Immunol. 2009 Aug 15;183(4):2767-74
references.pmid: 19625654
references.citation: J Periodontal Res. 1983 Mar;18(2):156-63
references.pmid: 6223995
references.citation: Proc Natl Acad Sci U S A. 1999 Jun 22;96(13):7220-5
references.pmid: 10377395
references.citation: Am J Physiol Cell Physiol. 2005 Nov;289(5):C1229-39
references.pmid: 15972838
references.citation: Biochem Biophys Res Commun. 1997 Jun 9;235(1):148-52
references.pmid: 9196052
references.citation: Biochem Biophys Res Commun. 1999 Nov 2;264(3):865-70
references.pmid: 10544022
references.citation: Arthritis Res Ther. 2004;6(4):R355-65
references.pmid: 15225371
references.citation: Arthritis Rheum. 2004 Feb;50(2):420-31
references.pmid: 14872484
references.citation: Mediators Inflamm. 2005 Oct 24;2005(5):249-55
references.pmid: 16258191
references.citation: J Pharmacol Exp Ther. 2005 Nov;315(2):788-95
references.pmid: 16081677
references.citation: J Periodontol. 2008 Mar;79(3):477-85
references.pmid: 18315430
references.citation: Proc Assoc Am Physicians. 1999 Nov-Dec;111(6):516-24
references.pmid: 10591080
references.citation: Mol Pharmacol. 2001 Mar;59(3):493-500
references.pmid: 11179444
references.citation: J Clin Invest. 2001 Jul;108(1):73-81
references.pmid: 11435459
references.citation: J Biol Chem. 2004 Mar 26;279(13):12647-58
references.pmid: 14722058
references.citation: J Periodontal Res. 1994 Mar;29(2):118-26
references.pmid: 8158500
references.citation: Int J Cancer. 2003 Nov 20;107(4):551-6
references.pmid: 14520691
references.citation: J Biol Chem. 2002 May 10;277(19):16355-64
references.pmid: 11847219
references.citation: Biochemistry. 2005 Jun 14;44(23):8326-36
references.pmid: 15938622
references.citation: J Immunol. 1998 Jan 1;160(1):403-9
references.pmid: 9551997
references.citation: Bioinformatics. 2007 Jan 15;23(2):257-8
references.pmid: 17098774
references.citation: J Periodontol. 1989 Sep;60(9):485-90
references.pmid: 2677301
references.citation: J Periodontol. 2009 May;80(5):833-49
references.pmid: 19405838
references.citation: J Periodontol. 2008 Sep;79(9):1645-51
references.pmid: 18771364
references.citation: Periodontol 2000. 1997 Jun;14:33-53
references.pmid: 9567965
references.citation: Quintessence Int. 2005 Apr;36(4):299-306
references.pmid: 15835427
references.citation: Int Immunopharmacol. 2006 Jun;6(6):987-96
references.pmid: 16644485
references.citation: Med Hypotheses. 2009 Sep;73(3):457-8
references.pmid: 19464122
references.citation: Clin Immunol. 2006 Jun;119(3):229-40
references.pmid: 16540375
references.citation: Bioinformatics. 2003 Feb 12;19(3):368-75
references.pmid: 12584122
references.citation: J Biol Chem. 2000 Oct 20;275(42):32775-82
references.pmid: 10922363
references.citation: Nat Genet. 2000 May;25(1):25-9
references.pmid: 10802651
references.citation: J Immunol. 2001 Jul 1;167(1):469-74
references.pmid: 11418684
references.citation: Proc Natl Acad Sci U S A. 2001 Nov 20;98(24):13681-6
references.pmid: 11717429
references.citation: J Biol Chem. 2000 Oct 20;275(42):32783-92
references.pmid: 10869354
references.citation: J Dent. 1998 Mar;26(2):97-103
references.pmid: 9540305
references.citation: Nucleic Acids Res. 2002 Feb 15;30(4):e15
references.pmid: 11842121
references.citation: Periodontol 2000. 1997 Jun;14:216-48
references.pmid: 9567973
references.citation: J Cell Physiol. 2003 Dec;197(3):348-56
references.pmid: 14566964
references.citation: Physiol Genomics. 2007 Oct 22;31(2):343-51
references.pmid: 17652167
references.citation: Arthritis Rheum. 2002 Mar;46(3):818-23
references.pmid: 11920420
references.citation: J Periodontal Res. 1991 Sep;26(5):415-21
references.pmid: 1832454
references.citation: Prostaglandins Other Lipid Mediat. 2002 Aug;68-69:115-28
references.pmid: 12432913
references.citation: J Clin Periodontol. 2002 Jan;29(1):15-20
references.pmid: 11846844
references.citation: Br J Pharmacol. 2008 Mar;153 Suppl 1:S241-6
references.pmid: 17965753
references.citation: Cell Physiol Biochem. 2006;17(1-2):69-78
references.pmid: 16543723
references.citation: BMC Med Genomics. 2008 Jul 07;1:30
references.pmid: 18606014
references.citation: Biochem Biophys Res Commun. 2002 Mar 8;291(4):884-9
references.pmid: 11866447
references.citation: J Periodontol. 2003 Mar;74(3):391-401
references.pmid: 12710761
references.citation: Periodontol 2000. 2007;43:85-101
references.pmid: 17214837
references.citation: Lancet. 2005 Nov 19;366(9499):1809-20
references.pmid: 16298220
references.citation: Am J Physiol Heart Circ Physiol. 2005 Jan;288(1):H165-74
references.pmid: 15358613
references.citation: Genome Biol. 2004;5(10):R80
references.pmid: 15461798
references.citation: J Pharmacol Exp Ther. 1999 Oct;291(1):124-30
references.pmid: 10490895
references.citation: Periodontol 2000. 1997 Jun;14:144-57
references.pmid: 9567969
references.citation: Eur J Oral Sci. 2004 Jun;112(3):216-23
references.pmid: 15154918
references.citation: J Biol Chem. 2006 Jun 16;281(24):16443-52
references.pmid: 16627484
references.citation: FEBS Lett. 2003 Jul 17;547(1-3):75-9
references.pmid: 12860389
references.citation: J Leukoc Biol. 2005 Mar;77(3):388-99
references.pmid: 15591425
references.citation: J Biol Chem. 2002 Jun 28;277(26):23573-81
references.pmid: 11950839
references.citation: Biochim Biophys Acta. 2007 Oct;1773(10):1589-98
references.pmid: 17707523
references.citation: J Dent Res. 1999 Jan;78(1):61-8
references.pmid: 10065947
references.citation: Cell Signal. 2006 Dec;18(12):2131-42
references.pmid: 16766159
references.citation: J Allergy Clin Immunol. 1978 Aug;62(2):96-102
references.pmid: 97337
references.citation: Int J Mol Med. 2006 Mar;17(3):449-55
references.pmid: 16465391
references.citation: Arthritis Res Ther. 2006;8(2):R47
references.pmid: 16507157
references.citation: J Clin Periodontol. 2005 Jan;32(1):6-11
references.pmid: 15642051
references.citation: Biochem Biophys Res Commun. 2003 Jun 13;305(4):970-3
references.pmid: 12767925
references.citation: Biostatistics. 2004 Jan;5(1):89-111
references.pmid: 14744830
references.citation: Sci STKE. 2006 Oct 17;2006(357):re13
references.pmid: 17047224
references.citation: Stat Appl Genet Mol Biol. 2004;3:Article3
references.pmid: 16646809
references.citation: Curr Drug Metab. 2007 Dec;8(8):750-7
references.pmid: 18220555
references.citation: Prostaglandins Other Lipid Mediat. 2002 Aug;68-69:383-99
references.pmid: 12432931
references.citation: Infect Immun. 1987 Sep;55(9):2148-54
references.pmid: 3040590
references.citation: BMC Genomics. 2005 Sep 28;6:137
references.pmid: 16191192
references.citation: J Immunol. 2001 Oct 1;167(7):3962-71
references.pmid: 11564815
references.citation: J Rheumatol. 2006 Dec;33(12):2398-408
references.pmid: 17225293
references.citation: Nucleic Acids Res. 2002 Jan 1;30(1):169-71
references.pmid: 11752283