12162497

pubmed:Citation

8

We tested the hypothesis that induction of cyclo-oxygenase (COX) 2 mediates some effects of bone morphogenetic protein (BMP) 2 on bone. BMP-2 induced COX-2 mRNA and prostaglandin (PG) production in cultured osteoblasts. BMP-2 increased luciferase activity in calvarial osteoblasts from mice transgenic for a COX-2 promoter-luciferase reporter construct (Pluc) and in MC3T3-E1 cells transfected with Pluc. Deletion analysis identified the -300/-213-bp region of the COX-2 promoter as necessary for BMP-2 stimulation of luciferase activity. Mutation of core-binding factor activity 1 (muCbfal) consensus sequence (5'-AACCACA3') at -267/-261 bp decreased BMP-2 stimulation of luciferase activity by 82%. Binding of nuclear proteins to an oligonucleotide spanning the Cbfal site was inhibited or supershifted by specific antibodies to Cbfal. In cultured osteoblasts from calvariae of COX-2 knockout (-/-) and wild-type (+/+) mice, the absence of COX-2 expression reduced the BMP-2 stimulation of both ALP activity and osteocalcin mRNA expression. In cultured marrow cells flushed from long bones, BMP-2 induced osteoclast formation in cells from COX-2(+/+) mice but not in cells from COX-2(-/-) mice. In vivo, BMP-2 (10 microg/pellet) induced mineralization in pellets of lyophilized collagen implanted in the flanks of mice. Mineralization of pellets, measured by microcomputed tomography (microCT), was decreased by 78% in COX-2(-/-) mice compared with COX-2(+/+) mice. We conclude that BMP-2 transcriptionally induces COX-2 in osteoblasts via a Cbfal binding site and that the BMP-2 induction of COX-2 can contribute to effects of BMP-2 on osteoblastic differentiation and osteoclast formation in vitro and to the BMP-2 stimulation of ectopic bone formation in vivo.

eng

IM

MEDLINE

0884-0431

Print

NLM

1430-40

pubmed-meshheading:12162497-3T3 Cells, pubmed-meshheading:12162497-Animals, pubmed-meshheading:12162497-Bone Morphogenetic Protein 2, pubmed-meshheading:12162497-Bone Morphogenetic Proteins, pubmed-meshheading:12162497-Calcification, Physiologic, pubmed-meshheading:12162497-Core Binding Factors, pubmed-meshheading:12162497-Cyclooxygenase 2, pubmed-meshheading:12162497-Dinoprostone, pubmed-meshheading:12162497-Electrophoretic Mobility Shift Assay, pubmed-meshheading:12162497-Enzyme Induction, pubmed-meshheading:12162497-Gene Expression Regulation, Enzymologic, pubmed-meshheading:12162497-Isoenzymes, pubmed-meshheading:12162497-Luciferases, pubmed-meshheading:12162497-Mice, pubmed-meshheading:12162497-Mutagenesis, Site-Directed, pubmed-meshheading:12162497-Neoplasm Proteins, pubmed-meshheading:12162497-Osteoblasts, pubmed-meshheading:12162497-Prostaglandin-Endoperoxide Synthases, pubmed-meshheading:12162497-RNA, Messenger, pubmed-meshheading:12162497-Transcription Factors, pubmed-meshheading:12162497-Transforming Growth Factor beta

Bone morphogenetic protein 2 induces cyclo-oxygenase 2 in osteoblasts via a Cbfal binding site: role in effects of bone morphogenetic protein 2 in vitro and in vivo.

Journal Article, In Vitro, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't