PMID- 12397613
OWN - NLM
STAT- MEDLINE
DCOM- 20040401
LR  - 20141120
IS  - 0730-2312 (Print)
IS  - 0730-2312 (Linking)
VI  - 87
IP  - 3
DP  - 2002
TI  - TNF-alpha suppresses bone sialoprotein (BSP) expression in ROS17/2.8 cells.
PG  - 313-23
AB  - Tumor necrosis factor-alpha (TNF-alpha) is a major mediator of inflammatory 
      responses in many diseases that inhibits bone formation and stimulates bone 
      resorption. To determine molecular mechanisms involved in the suppression of bone 
      formation we have analyzed the effects of TNF-alpha on BSP gene expression. Bone 
      sialoprotein (BSP) is a mineralized tissue-specific protein that appears to 
      function in the initial mineralization of bone. Previous studies have 
      demonstrated that BSP mRNA expression is essentially restricted to 
      fully-differentiated cells of mineralized connective tissues and that the 
      expression of BSP is developmentally regulated. Treatment of rat osteosarcoma ROS 
      17/2.8 cells with TNF-alpha (10 ng/ml) for 24 h caused a marked reduction in BSP 
      mRNA levels. The addition of antioxidant N-acetylcysteine (NAC; 20 mM) 30 min 
      prior to stimulation with TNF-alpha attenuated the inhibition of BSP mRNA levels. 
      Transient transfection analyses, using chimeric constructs of the rat BSP gene 
      promoter linked to a luciferase reporter gene, revealed that TNF-alpha (10 ng/ml) 
      suppressed expression in all constructs, including a short construct (pLUC3; nts 
      -116 to +60), transfected into ROS17/2.8 cells. Further deletion analysis of the 
      BSP promoter showed that a region within nts -84 to -60 was targeted by 
      TNF-alpha, the effects which were inhibited by NAC and the tyrosine kinase 
      inhibitor, herbimycin A (HA). Introduction of 2bp mutations in the inverted CCAAT 
      box (ATTGG; nts -50 and -46), a putative cAMP response element (CRE; nts -75 to 
      -68), and a FGF response element (FRE; nts -92 to -85) showed that the TNF-alpha 
      effects were mediated by the CRE. These results were supported by gel mobility 
      shift assays, using a radiolabeled double-stranded CRE oligonucleotide, which 
      revealed decreased binding of a nuclear protein from TNF-alpha-stimulated ROS 
      17/2.8 cells. Further, the inhibitory effect of TNF-alpha on CRE DNA-protein 
      complex was completely abolished by NAC or HA treatment. These studies, 
      therefore, show that TNF-alpha suppresses BSP gene transcription through a 
      tyrosine kinase-dependent pathway that generates reactive oxygen species and that 
      the TNF-alpha effects are mediated by a CRE element in the proximal BSP gene 
      promoter.
CI  - Copyright 2002 Wiley-Liss, Inc.
FAU - Samoto, Hiroshi
AU  - Samoto H
AD  - Department of Orthodontics, Nihon University School of Dentistry at Matsudo, 
      Chiba, Japan.
FAU - Shimizu, Emi
AU  - Shimizu E
FAU - Matsuda-Honjo, Yuko
AU  - Matsuda-Honjo Y
FAU - Saito, Ryoichiro
AU  - Saito R
FAU - Yamazaki, Muneyoshi
AU  - Yamazaki M
FAU - Kasai, Kazutaka
AU  - Kasai K
FAU - Furuyama, Shunsuke
AU  - Furuyama S
FAU - Sugiya, Hiroshi
AU  - Sugiya H
FAU - Sodek, Jaro
AU  - Sodek J
FAU - Ogata, Yorimasa
AU  - Ogata Y
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Cell Biochem
JT  - Journal of cellular biochemistry
JID - 8205768
RN  - 0 (Antioxidants)
RN  - 0 (Benzoquinones)
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (Ibsp protein, rat)
RN  - 0 (Integrin-Binding Sialoprotein)
RN  - 0 (Lactams, Macrocyclic)
RN  - 0 (Nuclear Proteins)
RN  - 0 (Quinones)
RN  - 0 (RNA, Messenger)
RN  - 0 (Sialoglycoproteins)
RN  - 0 (Tumor Necrosis Factor-alpha)
RN  - 1W306TDA6S (Rifabutin)
RN  - 70563-58-5 (herbimycin)
RN  - 9007-49-2 (DNA)
RN  - E0399OZS9N (Cyclic AMP)
RN  - EC 2.7.10.1 (Protein-Tyrosine Kinases)
RN  - WYQ7N0BPYC (Acetylcysteine)
SB  - IM
MH  - Acetylcysteine/pharmacology
MH  - Animals
MH  - Antioxidants/pharmacology
MH  - Benzoquinones
MH  - Cell Line
MH  - Cyclic AMP/genetics/metabolism
MH  - DNA/metabolism
MH  - Dose-Response Relationship, Drug
MH  - Down-Regulation/*drug effects
MH  - Electrophoretic Mobility Shift Assay/methods
MH  - Enzyme Inhibitors/pharmacology
MH  - Integrin-Binding Sialoprotein
MH  - Lactams, Macrocyclic
MH  - Nuclear Proteins/metabolism
MH  - Promoter Regions, Genetic/genetics
MH  - Protein-Tyrosine Kinases/antagonists & inhibitors/metabolism
MH  - Quinones/pharmacology
MH  - RNA, Messenger/biosynthesis
MH  - Rats
MH  - Response Elements/genetics
MH  - Rifabutin/analogs & derivatives
MH  - Sialoglycoproteins/antagonists & inhibitors/*biosynthesis/genetics
MH  - Transcription, Genetic/drug effects/physiology
MH  - Tumor Necrosis Factor-alpha/antagonists & inhibitors/*pharmacology
EDAT- 2002/10/25 04:00
MHDA- 2004/04/02 05:00
CRDT- 2002/10/25 04:00
PHST- 2002/10/25 04:00 [pubmed]
PHST- 2004/04/02 05:00 [medline]
PHST- 2002/10/25 04:00 [entrez]
AID - 10.1002/jcb.10301 [doi]
PST - ppublish
SO  - J Cell Biochem. 2002;87(3):313-23. doi: 10.1002/jcb.10301.