title: Regulation of osteoclastogenesis and RANK expression by TGF-beta1.
abstract: Transforming growth factor-beta (TGF-beta) has been shown to both inhibit and to stimulate bone resorption and osteoclastogenesis. This may be due, in part, to differential effects on bone marrow stromal cells that support osteoclastogenesis vs. direct effects on osteoclastic precursor cells. In the present study, we used the murine monocytic cell line, RAW 264.7, to define direct effects of TGF-beta on pre-osteoclastic cells. In the presence of macrophage-colony stimulating factor (M-CSF) (20 ng/ml) and receptor activator of NF-kappaB ligand (RANK-L) (50 ng/ml), TGF-beta1 (0.01-5 ng/ml) dose-dependently stimulated (by up to 120-fold) osteoclast formation (assessed by the presence of tartrate-resistant acid phosphatase (TRAP) positive multinucleated cells and expression of calcitonin and vitronectin receptors). In addition, TGF-beta1 also increased steady state RANK mRNA levels in a time- (by up to 3.5-fold at 48 h) and dose-dependent manner (by up to 2.2-fold at 10 ng/ml). TGF-beta1 induction of RANK mRNA levels was present both in undifferentiated RAW cells as well as in cells that had been induced to differentiate into osteoclasts by a 7-day treatment with M-CSF and RANK-L. Using a fluorescence-labeled RANK-L probe, we also demonstrated by flow cytometry that TGF-beta1 resulted in a significant increase in the percentage of RANK+ RAW cells (P < 0.05), as well as an increase in the fluorescence intensity per cell (P < 0.05), the latter consistent with an increase in RANK protein expression per cell. These data thus indicate that TGF-beta directly stimulates osteoclastic differentiation, and this is accompanied by increased RANK mRNA and protein expression.
authors.affiliation: Endocrine Research Unit, Mayo Clinic and Mayo Foundation, Rochester, Minnesota 55905, USA.
authors.forename: T
authors.initials: T
authors.lastname: Yan
authors.forename: B L
authors.initials: BL
authors.lastname: Riggs
authors.forename: W J
authors.initials: WJ
authors.lastname: Boyle
authors.forename: S
authors.initials: S
authors.lastname: Khosla
mesh_terms: D000135:Acid Phosphatase / Q000032:analysis / Q000378:metabolism; D000818:Animals; D015415:Biomarkers / Q000032:analysis; D002454:Cell Differentiation / Q000187:drug effects*; D006023:Glycoproteins / Q000819:agonists* / Q000096:biosynthesis; D007527:Isoenzymes / Q000032:analysis / Q000378:metabolism; D008264:Macrophages / Q000166:cytology; D051379:Mice; D010010:Osteoclasts / Q000166:cytology* / Q000378:metabolism*; D053244:Osteoprotegerin; D012333:RNA, Messenger / Q000819:agonists* / Q000235:genetics; D018003:Receptors, Calcitonin / Q000032:analysis / Q000378:metabolism; D018160:Receptors, Cytoplasmic and Nuclear / Q000819:agonists* / Q000096:biosynthesis; D018124:Receptors, Tumor Necrosis Factor; D019030:Receptors, Vitronectin / Q000032:analysis / Q000378:metabolism; D000071681:Tartrate-Resistant Acid Phosphatase; D016212:Transforming Growth Factor beta / Q000378:metabolism / Q000494:pharmacology*; D053773:Transforming Growth Factor beta1
chemical_list: D015415:Biomarkers; D006023:Glycoproteins; D007527:Isoenzymes; D053244:Osteoprotegerin; D012333:RNA, Messenger; D018003:Receptors, Calcitonin; D018160:Receptors, Cytoplasmic and Nuclear; D018124:Receptors, Tumor Necrosis Factor; D019030:Receptors, Vitronectin; C508961:Tgfb1 protein, mouse; C506031:Tnfrsf11b protein, mouse; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; D000135:Acid Phosphatase; C000605511:Acp5 protein, mouse; D000071681:Tartrate-Resistant Acid Phosphatase