title: 1,25-Dihydroxyvitamin D3 stimulates the production of insulin-like growth factor-binding proteins-2, -3 and -4 in human bone marrow stromal cells.
abstract: BACKGROUND
  1,25-Dihydroxyvitamin D3 (calcitriol) inhibits proliferation and stimulates differentiation of multiple cell types, including osteoblasts. Human (h) bone marrow stromal cells (MSCs) are a homogenous non-hematopoietic population of cells present in the bone marrow and exhibit a less differentiated osteoblastic phenotype. The IGF system, including IGFs-I, and -II and IGF binding proteins (IGFBPs), plays an important role in osteoblast cell proliferation and differentiation.
  
  
  OBJECTIVE
  To examine the pattern of expression of the IGF system in hMSCss and its regulation by calcitriol.
  
  
  RESULTS
  hMSCs express mRNA of both IGFs-I, and -II and IGFBPs-1 to -6 as shown by RT-PCR and northern blot analysis. As assessed by western ligand blotting (WLB) and western immmunoblot analysis, hMSCs secrete 38-42 kDa IGFBP-3, 24-28 kDa IGFBP-4 and a 33 kDa IGFBP-2. Calcitriol (dose range 10-10 mol/l) exerted no consistent dose-dependent effects on either IGF-I or IGF-II mRNA levels. In contrast, calcitriol treatment increased steady-state mRNA levels of IGFBPs-2, -3 and -4, but had no effect on IGFBP-5 or -6. Similarly, calcitriol increased the secretion of IGFBPs-2, -3 and -4 as determined by WLB. We found no detectable basal IGFBP-3 or IGFBP-4 protease activities in the absence or presence of calcitriol treatment.
  
  
  CONCLUSIONS
  Our results demonstrate that hMSCs expressed a distinct pattern of IGFs and IGFBPs that may be related to their stage of differentiation. The observed increase in production of IGFBPs-2, -3 and -4 by hMSCs upon treatment with calcitriol may be an important mechanism mediating the effects of calcitriol on MSC proliferation and differentiation.
authors.affiliation: Danish Center for Molecular Gerontology, , University of Aarhus, DK-8000 Aarhus C, Denmark.
authors.forename: M
authors.initials: M
authors.lastname: Kveiborg
authors.forename: A
authors.initials: A
authors.lastname: Flyvbjerg
authors.forename: E F
authors.initials: EF
authors.lastname: Eriksen
authors.forename: M
authors.initials: M
authors.lastname: Kassem
mesh_terms: D000328:Adult; D015152:Blotting, Northern; D015153:Blotting, Western; D001854:Bone Marrow Cells / Q000187:drug effects / Q000378:metabolism*; D002117:Calcitriol / Q000494:pharmacology*; D002478:Cells, Cultured; D017077:Culture Media, Conditioned; D010450:Endopeptidases / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D015151:Immunoblotting; D018971:Insulin-Like Growth Factor Binding Protein 2 / Q000096:biosynthesis*; D018972:Insulin-Like Growth Factor Binding Protein 3 / Q000096:biosynthesis*; D018974:Insulin-Like Growth Factor Binding Protein 4 / Q000096:biosynthesis*; D008666:Metalloendopeptidases / Q000378:metabolism; D011266:Pregnancy-Associated Plasma Protein-A; D012333:RNA, Messenger / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013268:Stimulation, Chemical; D017154:Stromal Cells / Q000187:drug effects / Q000378:metabolism; D013936:Thymidine / Q000378:metabolism
chemical_list: D017077:Culture Media, Conditioned; D018971:Insulin-Like Growth Factor Binding Protein 2; D018972:Insulin-Like Growth Factor Binding Protein 3; D018974:Insulin-Like Growth Factor Binding Protein 4; D012333:RNA, Messenger; D010450:Endopeptidases; D008666:Metalloendopeptidases; D011266:Pregnancy-Associated Plasma Protein-A; C080473:insulin-like growth factor binding protein-3 protease; D002117:Calcitriol; D013936:Thymidine