BMP-2 vs. BMP-4 expression and activity in glucocorticoid-arrested MC3T3-E1 osteoblasts: Smad signaling, not alkaline phosphatase activity, predicts rescue of mineralization

Growth Factors. 2008 Aug;26(4):226-37. doi: 10.1080/08977190802277880.

Abstract

Pharmacological glucocorticoids (GCs) inhibit bone formation, leading to osteoporosis. GCs inhibit bone morphogenetic protein-2 (Bmp2) expression, and rhBMP-2 restores mineralization in GC-arrested osteoblast cultures. To better understand how GCs regulate BMPs, we investigated Bmp transcription, as well as rhBMP-induced Smad and alkaline phosphatase (ALP) activity. Bmp2 cis-regulatory regions were analyzed by reporter plasmids and LacZ-containing bacterial artificial chromosomes. We found that GCs inhibited Bmp2 via a domain > 50 kb downstream of the coding sequence. Bmp expression was evaluated by RT-PCR; whereas GCs strongly inhibited Bmp2, Bmp4 was abundantly expressed and resistant to GCs. Both rhBMP-2 and rhBMP-4 restored mineralization in GC-arrested cultures; rhBMP-2 was 5-fold more effective when dosing was based on ALP activation, however, the rhBMPs were equipotent when dosing was based on Smad transactivation. In conclusion, GCs regulate Bmp2 via a far-downstream domain, and activation of Smad, not ALP, best predicts the pro-mineralization potential of rhBMPs.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3T3 Cells
  • Alkaline Phosphatase / analysis
  • Alkaline Phosphatase / metabolism
  • Animals
  • Bone Morphogenetic Protein 2 / genetics
  • Bone Morphogenetic Protein 2 / metabolism*
  • Bone Morphogenetic Protein 4 / genetics
  • Bone Morphogenetic Protein 4 / metabolism*
  • Calcification, Physiologic / drug effects*
  • Calcification, Physiologic / genetics
  • Gene Expression / drug effects*
  • Genes, Reporter
  • Glucocorticoids / pharmacology*
  • Humans
  • Luciferases / metabolism
  • Mice
  • Osteoblasts / drug effects*
  • Osteoblasts / metabolism
  • Recombinant Proteins / metabolism
  • Signal Transduction / drug effects
  • Transfection
  • beta-Galactosidase / analysis
  • beta-Galactosidase / metabolism

Substances

  • Bone Morphogenetic Protein 2
  • Bone Morphogenetic Protein 4
  • Glucocorticoids
  • Recombinant Proteins
  • Luciferases
  • Alkaline Phosphatase
  • beta-Galactosidase