Skip to main content
SpringerLink
Log in

Effects of Bone Morphogenetic Protein-2 and Transforming Growth Factor-β1 on Gene Expression of Decorin and Biglycan by Cultured Osteoblastic Cells

  • Published:
The Histochemical Journal Aims and scope Submit manuscript

Abstract

The influence of bone morphogenetic protein-2 (BMP-2) and transforming growth factor β (TGF-β) on the expression of small proteoglycans, decorin and biglycan was investigated in a clonal rat osteoblastic cell line, ROS-C26 (C26) cells, which is a potential osteoblast precursor cell line and capable of differentiating into mature osteoblasts after treatment with recombinant BMP-2 (rhBMP-2). Following the culture of C26 cells for 3, 6, and 9 days in the presence or absence of rhBMP-2, alkaline phosphatase activity increased in the rhBMP-2 treated cells in direct proportion to their differentiation into more mature osteoblastic cells, whereas decorin mRNA decreased in the cells, when compared to control cells without rhBMP-2 treatment. These results were evident 6 days after treatment. However, rhBMP-2 treatment had no effect on biglycan mRNA expression in the cells. Subsequently, after removal of rhBMP-2 from the culture media, the cells were further cultured for 24 h with graded concentrations of TGF-β1 (0, 0.1, 1.0, 5.0, and 10 ng/ml). TGF-β1 decreased decorin mRNA expression in the cells dose dependently, but did not affect their biglycan mRNA expression. Furthermore, either removal of rhBMP-2 from the culture media or addition of TGF-β1 significantly decreased alkaline phosphatase activity of rhBMP-2-induced cells. These results indicate that osteoblastic differentiation is accompanied by increased alkaline phosphatase activity and decreased expression of decorin mRNA, but continuous expression of biglycan mRNA. Both rhBMP-2 and TGF-β1 inhibit decorin mRNA expression in osteoblasts at varying stages of differentiation, but their effects on biglycan mRNA expression and alkaline phosphatase are different.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References cited

  • Abramson SR, Woessner JF Jr (1992) cDNA sequence for rat dermatan sulfate proteoglycan-II (decorin). Biochim Biophys Acta 1132: 225-227.

    Google Scholar 

  • Bianco P, Fisher LW, Young MF, Termine JD, Robey PG (1990) Expression and localization of the two small proteoglycans biglycan and decorin in developing human skeletal and non-skeletal tissues. J Histochem Cytochem 38: 1549-1563.

    Google Scholar 

  • Bianco P, Riminucci M, Fisher LW (1993) Biglycan and decorin in intact developing tissues: the in situ approach to their role in development, morphogenesis and tissue organization. In: Scott JE ed. Dermatan sulphate proteoglycans: Chemistry, Biology, Chemical Pathology. London: Portoland Press, pp. 193-205.

    Google Scholar 

  • Bossi M, Hoylaerts HF, Millan JL (1993) Modifications in a flexible surface loop modulate the isozyme-specific properties of mammalian alkaline phosphatases. J Biol Chem 268: 25409-25416.

    Google Scholar 

  • Breuer B, Schmidt G, Kresse H (1990) Non-uniform influence of transforming growth factor-β on the biosynthesis of different forms of small chondroitin sulphate/dermatan sulphate proteoglycan. Biochem J 269: 551-554.

    Google Scholar 

  • Centrella M, Horowitz MC, Wozney JM, Mccarthy TL (1994) Transforming growth factor-β gene family members and bone. Endocr Rev 15: 27-39.

    Google Scholar 

  • Chen TL, Mallory JB, Chang SL (1989) Modulation of transforming growth factor-β actions in rat osteoblast-like cells: the effects of bFGF and EGF. Growth Factors 1: 335-345.

    Google Scholar 

  • Chen TL, Bates RL, Dudley A, Hammonds RG Jr, Amento EP (1991) Bone morphogenetic protein-2b stimulation of growth and osteogenic phenotypes in rat osteoblast-like cells: comparison with TGF-β1. J Bone Miner Res 6: 1387-1393.

    Google Scholar 

  • Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidium thiocyanate-phenol-chloroform extraction. Anal Biochem 162: 156-159.

    Google Scholar 

  • Dreher KL, Asundi V, Matzura D, Cowan K (1990) Vascular smooth muscle biglycan represents a highly conserved proteoglycan within the arterial wall. Eur J Cell Biol 53: 296-304.

    Google Scholar 

  • Elford PR, Guenther HL, Felix R, Cecchini MG, Fleisch H (1987) Transforming growth factor-β reduces the phenotypic expression of osteoblastic MC3T3-E1 cells in monolayer culture. Bone 8: 259-262.

    Google Scholar 

  • Fisher LW, Termine JD, Dejter SW Jr, Whitson SW, Yanagishita M, Kimura JH, Hascall VC, Kleinman HK, Hassell JR, Nilsson B (1983) Proteoglycans of developing bone. J Biol Chem 258: 6588-6594.

    Google Scholar 

  • Fisher LW(1985) The nature of the proteoglycans of bone. In: Butler WT ed. The Chemistry and Biology of Mineralized Tissues. Birmingham: Ebsco Media, pp. 188-196.

    Google Scholar 

  • Fisher LW, Termine JD (1985) Noncollagenous proteins influencing the local mechanisms of calcification. Clin Orthop 200: 362-385.

    Google Scholar 

  • Fisher LW, Hawkins GR, Tuross N, Termine JD (1987) Purification and partial characterization of small proteoglycans I and II, bone sialoproteins I and II, and osteonectin from the mineral compartment of developing human bone. J Biol Chem 262: 9702-9708.

    Google Scholar 

  • Fisher LW, Termine JD, Young MF (1989) Deduced protein sequence of bone small proteoglycan I (biglycan) shows homology with proteoglycan II (decorin) and several nonconnective tissue proteins in a variety of species. J Biol Chem 264: 4571-4576.

    Google Scholar 

  • Franzén A, Heinegård D (1984a) Extraction and purification of proteoglycans from mature bovine bone. Biochem J 224: 47-58.

    Google Scholar 

  • Franzén A, Heinegård D (1984b) Characterization of proteoglycans from the calcified matrix of bovine bone. Biochem J 224: 59-66.

    Google Scholar 

  • Gunning P, Ponte P, Okayama H, Engel J, Blau H, Kedes L (1983) Isolation and characterization of full-length cDNAclones for human alpha-, beta-, and gamma-actin mRNAs: skeletal but not cytoplasmic actins have an amino-terminal cysteine that is subsequently removed. Mol Cell Biol 3: 787-795.

    Google Scholar 

  • Hedbom E, Heinegård D (1989) Interaction of a 59-kDa connective tissue matrix protein with collagen I and collagen II. J Biol Chem 264: 6898-6905.

    Google Scholar 

  • Heinegård D, Björne-Persson A, Cöster L, Franzén A, Gardell S, Malmström A, Paulsson M, Sandfalk R, Vogel K (1985) The core proteins of large and small interstitial proteoglycans from various connective tissues form distinct subgroups. Biochem J 230: 181-194.

    Google Scholar 

  • Hiraki Y, Inoue H, Shigeno C, Sanma Y, Bentz H, Rosen DM, Asada A, Suzuki F (1991) Bone morphogenetic proteins (BMP-2 and BMP-3) promote growth and expression of the differentiated phenotype of rabbit chondrocytes and osteoblastic MC3T3-E1 cells in vitro. J Bone Miner Res 6: 1373-1385.

    Google Scholar 

  • Hui MZ, Sukhu B, Tenenbaum HC (1996) Expression of tissue nonspecific alkaline phosphatase stimulates differentiated behaviour in specific transformed cell populations. Anat Rec 244: 423-436.

    Google Scholar 

  • Ibaraki K, Termine JD, Whitson SW, Young MF (1992) Bone matrix mRNA expression in differentiating fetal bovine osteoblasts. J Bone Miner Res 7: 743-754.

    Google Scholar 

  • Ibbotson KJ, Orcutt CM, Anglin AM, D'Souza SM (1989) Effects of transforming growth factors β1 and β2 on a mouse clonal, osteoblastlike cell line MC3T3-E1. J Bone Miner Res 4: 37-45.

    Google Scholar 

  • Katagiri T, Lee T, Takeshima H, Suda T, Tanaka H, Omura S (1990) Transforming growth factor-β modulates proliferation and differentiation of mouse clonal osteoblastic MC3T3-E1 cells depending on their maturation stages. Bone Miner 11: 285-293.

    Google Scholar 

  • Kimoto S, Cheng SL, Zhang SF, Avioli LV (1994) The effect of glucocorticoid on the synthesis of biglycan and decorin in human osteoblasts and bone marrow stromal cells. Endocrinology 135: 2423-2431.

    Google Scholar 

  • Lecanda F, AvioliL V, Cheng SL (1997) Regulation of bone matrix protein expression and induction of differentiation of human osteoblasts and human bone marrow stromal cells by bone morphogenetic protein-2. J Cell Biochem 67: 386-398.

    Google Scholar 

  • Noda M, Rodan GA (1986) Type-β transforming growth factor inhibits proliferation and expression of alkaline phosphatase in murine osteoblast-like cells. Biochem Biophys Res Commun 140: 56-65.

    Google Scholar 

  • Owen TA, Aronow M, Shalhoub V, Barone LM, Wilming L, Tassinari MS, Kennedy MB, Pockwinse S, Lian JB, Stein GS (1990) Progressive development of the rat osteoblast phenotype in vitro: reciprocal relationships in expression of genes associated with osteoblast proliferation and differentiation during formation of the bone extracellular matrix. J Cell Physiol 143: 420-430.

    Google Scholar 

  • Partridge NC, Alcorn D, Michelangeli VP, Kemp BE, Ryan GB, Martin TJ (1981) Functional properties of hormonally responsive cultured normal and malignant rat osteoblastic cells. Endocrinology 108: 213-219.

    Google Scholar 

  • Romaris M, Heredia A, Molist A, Bassols A (1991) Differential effect of transforming growth factor beta on proteoglycan synthesis in human embryonic lung fibroblasts. Biochim Biophys Acta 1093: 229-233.

    Google Scholar 

  • Rosen DM, Stempien SA, Thompson AY, Seyedin SM (1988) Transforming growth factor-beta modulates the expression of osteoblast and chondroblast phenotypes in vitro. J Cell Physiol 134: 337-346.

    Google Scholar 

  • Rosen V, Thies RS (1992) The BMP proteins in bone formation and repair. Trends Genet 8: 97-102.

    Google Scholar 

  • Ruoslahti E, Yamaguchi Y (1991) Proteoglycans as modulators of growth factor activities. Cell 64: 867-869.

    Google Scholar 

  • Thiebaud D, Guenther HL, Porret A, Burckhardt P, Fleisch H, Hofstetter W (1994) Regulation of collagen type I and biglycan mRNA levels by hormones and growth factors in normal and immortalized osteoblastic cell lines. J Bone Miner Res 9: 1347-1354.

    Google Scholar 

  • Takeuchi Y, Matsumoto T, Ogata E, Shishiba Y (1993) Effects of transforming growth factor β1 and L-ascorbate on synthesis and distribution of proteoglycans in murine osteoblast-like cells. J Bone Miner Res 8: 823-830.

    Google Scholar 

  • Takeuchi Y, Fukumoto S, Matsumoto T (1995) Relationship between actions of transforming growth factor (TGF)-β and cell surface expression of its receptors in clonal osteoblastic cells. J Cell Physiol 162: 315-321.

    Google Scholar 

  • Vogel KG, Paulsson M, Heinegård D (1984) Specific inhibition of type I and type II collagen fibrillogenesis by the small proteoglycan of tendon. Biochem J 223: 587-597.

    Google Scholar 

  • Westergren-Thorsson G, Antonsson P, Malmström A, Heinegård D, Oldberg Å. (1991) The synthesis of a family of structurally related proteoglycans in fibroblasts is differently regulated by TGF-β. Matrix 11: 177-183.

    Google Scholar 

  • Xu T, Bianco P, Fisher LW, Longenecker G, Smith E, Goldstein S, Bonadio J, Boskey A, Heegaard AM, Sommer B, Satomura K, Dominguez P, Zhao C, Kulkarni AB, Robey PG, Young MF (1998) Targeted disruption of the biglycan gene leads to an osteoporosis-like phenotype in mice. Nat Genet 20: 78-82.

    Google Scholar 

  • Yamaguchi Y, Mann DM, Ruoslahti E (1990) Negative regulation of transforming growth factor-β by the proteoglycan decorin. Nature 346: 281-284.

    Google Scholar 

  • Yamaguchi A, Katagiri T, Ikeda T, Wozney JM, Rosen V, Wang EA, Kahn AJ, Suda T, Yoshiki S (1991) Recombinant human bone morphogenetic protein-2 stimulates osteoblastic maturation and inhibits myogenic differentiation in vitro. J Cell Biol 113: 681-687.

    Google Scholar 

  • Yamaguchi A, Kahn AJ (1991) Clonal osteogenic cell lines express myogenic and adipocytic developmental potential. Calcif Tissue Int 49: 221-225.

    Google Scholar 

  • Yanagishita M (1993) Function of proteoglycans in the extracellular matrix. Acta Pathol Jpn 43: 283-293.

    Google Scholar 

  • Yoon K, Golub E, Rodan GA (1989) Alkaline phosphatase cDNA transfected cells promote calcium and phosphate deposition. Connect Tissue Res 22: 17-25.

    Google Scholar 

  • Zhou H, Hammonds RG Jr, Findlay DM, Fuller PJ, Martin TJ, Ng KW (1991) Retinoic acid modulation of mRNA levels in malignant, nontransformed, and immortalized osteoblasts. J Bone Miner Res 6: 767-777.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Anatomy, Nihon University School of Dentistry, 1-8-13 Kanda-Surugadai, Chiyoda-ku, Tokyo, 101-8310, Japan

    Minoru Takagi, Toshihiro Yamada, Naoko Kamiya & Tohru Kumagai

  2. Department of Oral Pathology, Nagasaki University School of Dentistry, Nagasaki, 852-8588, Japan

    Akira Yamaguchi

Authors
  1. Minoru Takagi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Toshihiro Yamada
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Naoko Kamiya
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Tohru Kumagai
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Akira Yamaguchi
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Takagi, M., Yamada, T., Kamiya, N. et al. Effects of Bone Morphogenetic Protein-2 and Transforming Growth Factor-β1 on Gene Expression of Decorin and Biglycan by Cultured Osteoblastic Cells. Histochem J 31, 403–409 (1999). https://doi.org/10.1023/A:1003704425809

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1003704425809

Keywords

Search

Navigation