title: Role of NFkappaB in the regulation of macrophage colony stimulating factor by tumor necrosis factor-alpha in ST2 bone stromal cells.
abstract: Expression of MCSF in bone is important to the regulation of osteoclastogenesis. We show here that tumor necrosis factor-alpha (TNFalpha) increases the production of both soluble (sMCSF) and membrane-bound (mMCSF) macrophage colony stimulating factor by ST2 bone stromal cells. Treatment of ST2 cells with TNFalpha caused sMCSF levels to increase by 394+/-5% from basal; mMCSF rose by 316+/-66% from 30+/-10 per 100,000 cells in the same time. These increases were consistent with increased expression of mRNAs encoding both isoforms. Increases in MCSF mRNA are also seen after stimulation with dexamethasone. To investigate the potential role of NFkappaB in this TNFalpha effect, we treated cells with sodium salicylate (NaS), an inhibitor of NFkappaB translocation. NaS decreased TNFalpha-stimulated NFkappaB activation by 50% as assessed by EMSA. Despite inhibition of NFkappaB signaling, NaS enhanced TNFalpha-stimulated MCSF secretion and did not prevent TNFalpha-stimulated increases in sMCSF mRNA, suggesting that NFkappaB was not involved in TNFalpha effect on the gene. TNFalpha failed to stimulate transcription of a 774 nucleotide MCSF promoter-luciferase reporter transfected into ST2 cells which contained the NFkappaB consensus sequence. Deletion of the seven nucleotides containing the NFkappaB homology response sequence from the MCSF promoter increased basal gene transcription by twofold. TNFalpha thus contributes to an osteoclastogenic environment through upregulation of bone expression of both MCSF isoforms. Our data suggests that NFkappaB is not the major signaling pathway through which this occurs.
authors: Department of Medicine, Emory University and Veterans Affairs Medical Center, Atlanta, Georgia 30033, USA.
authors: S D
authors: SD
authors: Isaacs
authors: X
authors: X
authors: Fan
authors: D
authors: D
authors: Fan
authors: H
authors: H
authors: Gewant
authors: T C
authors: TC
authors: Murphy
authors: P
authors: P
authors: Farmer
authors: W R
authors: WR
authors: Taylor
authors: M S
authors: MS
authors: Nanes
authors: J
authors: J
authors: Rubin
mesh_terms: D001842:Bone and Bones / Q000187:drug effects / Q000378:metabolism*; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000032:analysis; D003907:Dexamethasone / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D017930:Genes, Reporter; D016173:Macrophage Colony-Stimulating Factor / Q000378:metabolism*; D016328:NF-kappa B / Q000378:metabolism*; D009687:Nuclear Proteins / Q000032:analysis; D012333:RNA, Messenger / Q000378:metabolism; D012980:Sodium Salicylate / Q000494:pharmacology; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
chemical_list: D004268:DNA-Binding Proteins; D016328:NF-kappa B; D009687:Nuclear Proteins; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D003907:Dexamethasone; D016173:Macrophage Colony-Stimulating Factor; D012980:Sodium Salicylate