PMID- 15516323
OWN - NLM
STAT- MEDLINE
DCOM- 20050324
LR  - 20061115
IS  - 1065-6995 (Print)
IS  - 1065-6995 (Linking)
VI  - 28
IP  - 10
DP  - 2004
TI  - Effect of TNF-alpha on human osteosarcoma cell line Saos2--TNF-alpha regulation 
      of bone sialoprotein gene expression in Saos2 osteoblast-like cells.
PG  - 653-60
AB  - Tumor necrosis factor-alpha (TNF-alpha) is a major mediator of inflammatory 
      response in many diseases. It inhibits bone formation and stimulates bone 
      resorption. To determine the molecular mechanisms involved in the regulation of 
      gene expression of osteoblast-like cells, we analyzed the effects of TNF-alpha on 
      the human osteosarcoma cell line Saos2. We used RT-PCR to examine the effects of 
      TNF-alpha on bone sialoprotein (BSP), core binding factor a1 (Cbfa1), osterix, 
      alpha 1 (I) collagen, cyclooxygenase-2 (COX-2), interleukin-6 (IL-6), cathepsin 
      B, cathepsin L and tissue inhibitors of metalloproteinase-1 (TIMP-1). TNF-alpha 
      (10ng/ml) increased BSP, IL-6 and COX-2 mRNA levels after 3h, reaching maximal 
      levels at 12 h. Cbfa1 mRNA levels increased after 3 h, but decreased by 24 h. 
      Osterix, cathepsin B, cathepsin L and TIMP-1 mRNA levels did not change after 
      stimulation with TNF-alpha. On the other hand, alpha 1 (I) collagen mRNA 
      expression was suppressed by TNF-alpha at 24 h. Transient transfection analyses 
      were performed using chimeric constructs of the rat BSP gene promoter linked to a 
      luciferase reporter gene. TNF-alpha (10 ng/ml) had no effect on the promoter 
      activities of BSP transfected into Saos2 cells. The results of gel mobility shift 
      assays using radiolabeled double-stranded cAMP response element (CRE) and FGF2 
      response element (FRE) oligonucleotides in the proximal promoter of the rat BSP 
      gene showed increased binding of nuclear proteins at 6 h. Gel mobility shift 
      assays with radiolabelled COX-2-CRE and COX-2-NF kappa B oligonucleotides 
      revealed an increase in the binding of nuclear proteins from TNF-alpha-stimulated 
      Saos2 cells. These studies, therefore, showed that TNF-alpha indirectly increased 
      BSP expression, and that it could be mediated through COX-2 and Cbfa1 expression 
      in Saos2 osteoblast-like cells.
FAU - Nakayama, Youhei
AU  - Nakayama Y
AD  - Department of Periodontology, Nihon University School of Dentistry at Matsudo, 
      Chiba 271-8587, Japan.
FAU - Kato, Naoko
AU  - Kato N
FAU - Nakajima, Yu
AU  - Nakajima Y
FAU - Shimizu, Emi
AU  - Shimizu E
FAU - Ogata, Yorimasa
AU  - Ogata Y
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Cell Biol Int
JT  - Cell biology international
JID - 9307129
RN  - 0 (Core Binding Factor Alpha 1 Subunit)
RN  - 0 (Core Binding Factors)
RN  - 0 (Membrane Proteins)
RN  - 0 (Neoplasm Proteins)
RN  - 0 (RNA, Messenger)
RN  - 0 (SPP1 protein, human)
RN  - 0 (Sialoglycoproteins)
RN  - 0 (Spp1 protein, rat)
RN  - 0 (Transcription Factors)
RN  - 0 (Tumor Necrosis Factor-alpha)
RN  - 106441-73-0 (Osteopontin)
RN  - EC 1.14.99.1 (Cyclooxygenase 2)
RN  - EC 1.14.99.1 (PTGS2 protein, human)
RN  - EC 1.14.99.1 (Prostaglandin-Endoperoxide Synthases)
SB  - IM
MH  - Cell Line, Tumor
MH  - Core Binding Factor Alpha 1 Subunit
MH  - Core Binding Factors
MH  - Cyclooxygenase 2
MH  - Gene Expression Regulation, Neoplastic/physiology
MH  - Humans
MH  - Membrane Proteins
MH  - Neoplasm Proteins/biosynthesis/genetics/physiology
MH  - Osteoblasts/*metabolism
MH  - Osteopontin
MH  - Osteosarcoma/genetics/*metabolism
MH  - Prostaglandin-Endoperoxide Synthases/biosynthesis/genetics/physiology
MH  - RNA, Messenger/biosynthesis/genetics
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Sialoglycoproteins/*biosynthesis/genetics
MH  - Transcription Factors/biosynthesis/genetics/physiology
MH  - Transfection
MH  - Tumor Necrosis Factor-alpha/*physiology
EDAT- 2004/11/02 09:00
MHDA- 2005/03/25 09:00
CRDT- 2004/11/02 09:00
PHST- 2004/02/27 00:00 [received]
PHST- 2004/05/06 00:00 [revised]
PHST- 2004/06/03 00:00 [accepted]
PHST- 2004/11/02 09:00 [pubmed]
PHST- 2005/03/25 09:00 [medline]
PHST- 2004/11/02 09:00 [entrez]
AID - S1065-6995(04)00134-9 [pii]
AID - 10.1016/j.cellbi.2004.06.003 [doi]
PST - ppublish
SO  - Cell Biol Int. 2004;28(10):653-60. doi: 10.1016/j.cellbi.2004.06.003.