title: Mouse embryo-derived NIH3T3 fibroblasts adopt an osteoblast-like phenotype when treated with 1alpha,25-dihydroxyvitamin D(3) and dexamethasone in vitro.
abstract: This study examines the capability of NIH3T3 fibroblasts to express osteoblastic markers following stimulation with a number of hormones and growth factors in vitro. Of the agents tested, 1alpha,25-dihydroxyvitamin D(3) (1,25(OH)(2)D(3)) dose-dependently induced alkaline phosphatase (ALP) activity in NIH3T3 cells, and this effect was enhanced by the addition of dexamethasone (Dex), which when administered alone caused no detectable ALP expression. The combined use of 1,25(OH)(2)D(3) and Dex also stimulated the synthesis of osteocalcin, and osteopontin. Furthermore, cells treated with the both hormones, in the presence of beta-glycerophosphate and l-ascorbic acid, formed mineralized plaques, indicating an osteoblast (OB) phenotype. By contrast, the differentiation induced by 1,25(OH)(2)D(3) or 1,25(OH)(2)D(3) plus Dex was significantly antagonized by transforming growth factor-beta1 and all trans-retinoic acid. These data indicate that NIH3T3 cells have the potential to adopt an OB-like phenotype and may prove to be a convenient model for studying the early events of osteogenic differentiation and the specific interactions of 1,25(OH)(2)D(3) with glucocorticoids in controlling this process in vitro.
authors.affiliation: Department of Human Metabolism and Clinical Biochemistry, University of Sheffield Medical School, Sheffield, United Kingdom.
authors.forename: Chaoxiang
authors.initials: C
authors.lastname: Shui
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Scutt
mesh_terms: D016475:3T3 Cells; D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D015415:Biomarkers; D002113:Calcification, Physiologic; D002117:Calcitriol / Q000008:administration & dosage / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D003094:Collagen / Q000378:metabolism; D003907:Dexamethasone / Q000008:administration & dosage / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology; D005994:Glycerophosphates / Q000378:metabolism; D051379:Mice; D009363:Neoplasm Proteins; D010006:Osteoblasts / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D015675:Osteocalcin / Q000378:metabolism; D053495:Osteopontin; D010641:Phenotype; D012795:Sialoglycoproteins / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000494:pharmacology; D053773:Transforming Growth Factor beta1; D014212:Tretinoin / Q000494:pharmacology
chemical_list: D015415:Biomarkers; D005994:Glycerophosphates; D009363:Neoplasm Proteins; D012795:Sialoglycoproteins; C506896:Spp1 protein, mouse; C508961:Tgfb1 protein, mouse; D014157:Transcription Factors; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; D015675:Osteocalcin; D053495:Osteopontin; D014212:Tretinoin; D003907:Dexamethasone; D003094:Collagen; D000469:Alkaline Phosphatase; D002117:Calcitriol; C031463:beta-glycerophosphoric acid