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Endocrinology Vol. 141, No. 8 2914-2922
Copyright © 2000 by The Endocrine Society

ARTICLES

Nuclear Factor-{kappa}B p50 Is Required for Tumor Necrosis Factor-{alpha}-Induced Colony-Stimulating Factor-1 Gene Expression in Osteoblasts1

Gang-Qing Yao, Ben-hua Sun, Karl L. Insogna2 and Eleanor C. Weir2,3

The Section of Comparative Medicine (G.-Q.Y., E.C.W.), and the Department of Internal Medicine (B.-H.S., K.L.I.), Yale University School of Medicine, New Haven, Connecticut 06520

Address all correspondence and requests for reprints to: Gang-Qing Yao, Section of Comparative Medicine, Yale University School of Medicine, P.O. Box 208016, New Haven, Connecticut 06520-8016. E-mail: gang-qing.yao{at}yale.edu

Colony-stimulating factor (CSF)-1 is a hematopoietic growth factor that is released by osteoblasts and is recognized to play a critical role in bone remodeling in vivo and in vitro. We have reported that osteoblasts express CSF-1 constitutively and that tumor necrosis factor (TNF)-{alpha}, a potent bone-resorbing agent, increases CSF-1 gene expression by a transcriptional mechanism. In the present study, we report that an NF-{kappa}B site in the CSF-1 promoter is required for TNF-{alpha}-induced CSF-1 expression in osteoblasts. As determined by electrophoretic mobility shift assays, antiserum against the NF-{kappa}B-binding protein, p50, retarded the mobility of the inducible complex, whereas antisera against p52, p65, c-Rel, Rel B, I{kappa}B {alpha}, I{kappa}B {gamma}, and Bcl-3 had no effect. To further confirm that p50 is necessary for TNF-{alpha}-induced CSF-1 expression in osteoblasts, CSF-1 messenger RNA expression from untreated and TNF-{alpha}-treated osteoblasts, prepared from wild-type and p50 knock-out mice, was examined by Northern analysis. CSF-1 messenger RNA was increased by TNF treatment in wild-type mice but not in NF-{kappa}B p50 knock-out mice. Our findings support the conclusion that the NF-{kappa}B subunit p50 is critical for TNF-induced CSF-1 expression in osteoblasts.




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