Differentiation and cell surface expression of transforming growth factor-beta receptors are regulated by interaction with matrix collagen in murine osteoblastic cells

J Biol Chem. 1996 Feb 16;271(7):3938-44. doi: 10.1074/jbc.271.7.3938.


Although transforming growth factor (TGF)-beta enhances bone formation, it inhibits the differentiation of osteoblasts. To clarify the regulatory mechanism of osteoblastic differentiation and TGF-beta actions, the relationship among differentiation, TGF-beta actions, and matrix protein synthesis was examined using murine osteoblast-like MC3T3-E1 cells. Alkaline phosphatase (ALP) activity continued to increase during long-term cultures, and the increase was closely associated with a reduction in cell surface TGF-beta receptors competent to bind TGF-beta. Both the stimulation of proteoglycan synthesis and the inhibition of ALP activity by TGF-beta were also suppressed. Collagen synthesis inhibitors and an anti-alpha2beta1 integrin blocking antibody blocked the changes in ALP activity and TGF-beta receptors, and a DGEA peptide that interferes binding of collagen to alpha2beta1 integrin also blocked the increase in ALP activity. Furthermore, when MC3T3-E1 cells were cultured on extracellular matrix layers obtained from these cells, all the differentiation-associated changes could be observed without collagen production, and the extracellular matrix-induced differentiation was also blocked by an anti-alpha2beta1 integrin antibody. These results demonstrate that the interaction of cell surface alpha2beta1 integrin with matrix collagen synthesized by osteoblasts themselves is involved in the osteoblastic differentiation and the reduction in cell surface receptors and actions of TGF-beta. It is suggested that matrix collagen synthesized under the stimulation by TGF-beta plays an important role in the regulation of osteoblastic differentiation and TGF-beta actions by differentiation-associated down-regulation of TGF-beta receptors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3T3 Cells
  • Alkaline Phosphatase / biosynthesis
  • Amino Acid Sequence
  • Animals
  • Ascorbic Acid / pharmacology
  • Azetidinecarboxylic Acid / pharmacology
  • Blotting, Northern
  • Cell Differentiation*
  • Cell Membrane / metabolism
  • Collagen / biosynthesis
  • Collagen / physiology*
  • Extracellular Matrix*
  • Gene Expression Regulation*
  • Integrins / antagonists & inhibitors
  • Integrins / physiology
  • Mice
  • Molecular Sequence Data
  • Oligopeptides / pharmacology
  • Osteoblasts / cytology*
  • Osteoblasts / metabolism*
  • Proline / pharmacology
  • Proteoglycans / biosynthesis
  • Receptors, Collagen
  • Receptors, Transforming Growth Factor beta / biosynthesis*


  • Integrins
  • Oligopeptides
  • Proteoglycans
  • Receptors, Collagen
  • Receptors, Transforming Growth Factor beta
  • Azetidinecarboxylic Acid
  • Collagen
  • Proline
  • Alkaline Phosphatase
  • Ascorbic Acid