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1,25-Dihydroxyvitamin D3 metabolism in a human osteosarcoma cell line and human bone cells.

Abstract
The metabolism of 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] by a human osteoblastic sarcoma cell line, U-2 OS, and by primary cultures of human bone-derived cells was examined at physiologic (5 x 10(-11) M) and pharmacologic (3.5 x 10(-7) M) substrate concentrations. For metabolite identification purposes, cells nearing confluency were incubated for 18 h with 3.5 x 10(-7) M 1,25-(OH)2D3 in serum-free medium. The putative vitamin D metabolites produced during this incubation were isolated from a total lipid extract of cells and medium. Identification of the metabolites was achieved by comigration with authentic standards on three high-performance liquid chromatography systems, UV spectral analysis, mass spectrometry, and chemical modification by sodium borohydride and sodium metaperiodate. The identified metabolites produced from 1,25-(OH)2D3 by the human osteosarcoma cells include 1,24,25-trihydroxyvitamin D3; 24-oxo-1,25-dihydroxyvitamin D3; 24-oxo-1,23,25-trihydroxyvitamin D3; and 24,25,26,27-tetranor-1,23-dihydroxyvitamin D3. Evidence is presented that (1) 1,25-(OH)2D3 metabolism occurs constitutively in U-2 OS osteosarcoma cells at a physiologic substrate concentration (5 x 11(-11) M), (2) the pathway can be further induced by pharmacologic 1,25-(OH)2D3 concentrations (10(-7) M), and (3) this pathway is present in primary cultures of normal human bone-derived cells.
AuthorsB E Miller, D P Chin, G Jones
JournalJournal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research (J Bone Miner Res) Vol. 5 Issue 6 Pg. 597-608 (Jun 1990) ISSN: 0884-0431 [Print] United States
PMID2166423 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • Lipids
  • Trimethylsilyl Compounds
  • Periodic Acid
  • metaperiodate
  • Calcitriol
Topics
  • Bone and Bones (cytology, metabolism)
  • Calcitriol (metabolism)
  • Cells, Cultured
  • Chromatography, High Pressure Liquid
  • Humans
  • Lipids (isolation & purification)
  • Mass Spectrometry
  • Osteosarcoma (metabolism)
  • Periodic Acid
  • Trimethylsilyl Compounds
  • Tumor Cells, Cultured

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