Skip to main content
SpringerLink
Log in

Modulation of proteases and their inhibitors in immortal human osteoblast-like cells by tumor necrosis factor-alpha in vitro

  • Original Articles
  • Published:
Inflammation Aims and scope Submit manuscript

Abstract

Inflammatory cytokines like interleukin-1 (IL-1) and tumor necrosis factor-alpha (TNF-α) are linked to abnormal cartilage and bone loss in a variety of pathological conditions. We have investigated the effect of TNF-α on the synthesis and/ or steady-state mRNA levels of collagen, alkaline phosphatase (ALP), plasminogen activators (PAs) and their inhibitor PAI-1, and collagenases (MMPs) and their inhibitor TIMP-1 by human osteoblastic, HOS TE85, cells in monolayer cultures. HOS TE85 cells possess ∼2000 TNF-α receptors per cell with aK d value of 0.67 nM and receptor of ∼ 60 kDa. TNF-α enhances urokinase-plasminogen activator (u-PA) activity and steady-state mRNA levels twofold without affecting tissue-plasminogen activator (t-PA) or PAI-1. The increase in u-PA mRNA is due to enhanced transcription of this gene. mRNA levels or activities of collagenase 1 (MMP-1), 72- and 92-kDa gelatinases (MMP-2 and MMP-9) are also nearly doubled with little change in the level of expression of TIMP-1. TNF-α does not significantly affect the activity or mRNA levels of ALP. TNF-α decreases collagen as well as general protein synthesis. However, the steady-state mRNA for the α2 chain of collagen type I is increased three- to fourfold. These results show that TNF-α may increase pathological bone turnover by enhancing the rate of transcription of proteases capable of degrading the nonmineralized osteoid layer and decelerating the maturation of the extracellular matrix formed by osteoblasts.

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

  1. Le, J., andJ. Vilcek. 1987. Tumor necrosis factor and Interleukin 1: Cytokines with multiple overlapping biological activities.Lab. Invest. 56:234–248.

    PubMed  Google Scholar 

  2. Camussi, G., E. Albano, C. Tetta, andF. Bussolino. 1991. The molecular action of tumor necrosis factor-α.Eur. J. Biochem. 202:3–14.

    PubMed  Google Scholar 

  3. Rosenblum, M. G., andN. J. Donato. 1989. Tumor necrosis factor α: A multifaceted peptide hormone.Crit. Rev. Immunol. 9:21–44.

    PubMed  Google Scholar 

  4. Hohmann, H.-P., R. Remy, M. Brockhaus, andA. P. Van Loon. 1989. Two different cells types have different major receptors for human tumor necrosis factor.J. Biol. Chem. 264:14927–14934.

    PubMed  Google Scholar 

  5. Brockhaus, M., H. J. Schoenfeld, E. J. Schlaeger, W. Hunziker, W. Leslauer, andH. Loetscher. 1990. Identification of two types of tumor necrosis factor receptors on human cell lines by monoclonal antibodies.Proc. Natl. Acad. Sci. U. S. A. 87:3127–3131.

    PubMed  Google Scholar 

  6. Alvaro-Garcia, J. M., N. J. Zvaifler, C. B. Brown, K. Kaushansky, andG. S. Firestein. 1991. Cytokines in chronic inflammatory arthrities. IV. Analysis of the synovial cells involved in granulocyte-macrophage colony-stimulating factor production and gene expression in rheumatoid arthritis and its regulation by IL-1 and tumor necrosis factor-α.J. Immunol. 146:3365–3371.

    PubMed  Google Scholar 

  7. Sugarman, B. J., B. B. Aggarwal, P. E. Hass, I. S. Figari, M. A. Palladino, Jr., andH. M. Shepard. 1985. Recombinant human tumor necrosis factor-α: Effects of proliferation of normal and transformed cells in vitro.Science 230:943–945.

    PubMed  Google Scholar 

  8. Lewis, G. D., B. B. Aggarwal, T. E. Eessalu, B. J. Sugarman, andH. M. Shepard. 1987. Modulation of the growth of transformed cells by human tumor necrosis factor-α and interferon-γ.Cancer Res. 47:5382–5385.

    PubMed  Google Scholar 

  9. Bertolini, D. R., G. E. Nedwin, T. S. Bringman, D. D. Smith, andG. R. Mundy. 1986. Stimulation of bone resorption and inhibition of bone formation in vitro by human tumor necrosis factor.Nature 319:516–518.

    PubMed  Google Scholar 

  10. Akira, S., T. Hirano, T. Taga, andT. Kishimoto. 1990. Biology of multifunctional cytokines: IL-6 and related molecules (IL-1 and TNF).FASEB J. 4:2860–2867.

    PubMed  Google Scholar 

  11. Rossomando, E. F., J. E. Kennedy, andJ. Hadjimichael. 1990. Tumor necrosis factor alpha in gingival crevicular fluid as a possible indicator of periodontal disease in humans.Arch. Oral Biol. 35:431–434.

    PubMed  Google Scholar 

  12. Alvaro-Garcia, J. M., N. J. Zvaifler, andG. S. Firestein. 1990. Cytokines in chronic inflammatory arthritis. V. Mutual antagonism between interferon-gamma and tumor necrosis factor-alpha on HLA-DR expression, proliferation, collagenase production and granulocyte macrophage colony-stimulating factor production by rheumatoid arthritis synoviocytes.J. Clin. Invest. 86:1790–1798.

    PubMed  Google Scholar 

  13. Duncan, M. R., andB. Berman. 1989. Differential regulation of collagen, glycosaminoglycan, fibronectin, and collagenase activity in cultured human adult dermal fibroblasts by interleukin 1-alpha and beta and tumor necrosis factor-alpha and beta.J. Invest. Dermatol. 92:699–706.

    PubMed  Google Scholar 

  14. MacNaul, K. L., N. Chartrain, M. Lark, M. J. Tocci, andN. I. Hutchinson. 1990. Discoordinate expression of stromelysin, collagenase, and tissue inhibitor of metalloproteinases-1 in rheumatoid synovial fibroblasts.J. Biol. Chem. 265:17238–17245.

    PubMed  Google Scholar 

  15. Lefebvre, V., C. Peeters-Joris, andG. Vaes. 1991. Production of gelatin-degrading matrix metalloproteinases (‘type IV collagenases’) and inhibitors by articular chondrocytes during their dedifferentiation by serial subcultures and under stimulation by interleukin-1 and tumor necrosis factor-α.Biochim. Biophys. Acta 1094:8–18.

    PubMed  Google Scholar 

  16. Campbell, I. K., D. S. Piccoli, M. J. Roberts, K. D. Muirden, andJ. A. Hamilton. 1990. Effects of tumor necrosis factor alpha and beta on resorption of human articular cartilage and production of plasminogen activator by human articular chondrocytes.Arthritis Rheum. 33:543–552.

    Google Scholar 

  17. Canalis, E. 1987. Effects of tumor necrosis factor on bone formation in vitro.Endocrinology 121:1596–1604.

    PubMed  Google Scholar 

  18. Panagakos, F. S., L. Hinojosa, andS. Kumar. 1992. TNF modulation of bone formation and bone resorption in vitro.J. Dent. Res. 71:149.

    Google Scholar 

  19. Jones, P. A., J. S. Rhim, H. Issacs, andR. M. McAlister. 1975. Relationship between tumorigenicity, growth in agar, and fibrinolytic activity in a line of human osteosarcoma cells.Int. J. Cancer 16:616–621.

    PubMed  Google Scholar 

  20. Ding, A. H., E. Sanchez, S. Srimal, C. F. Nathan 1989. Macrophages rapidly internalize their tumor necrosis factor receptors in response to bacterial lipopolysaccharide.J. Biol. Chem. 264:3924–3929.

    PubMed  Google Scholar 

  21. Smith, R. A., andC. Baglioni. 1989. Multimeric structure of the tumor necrosis factor receptor of HeLa cells.X Biol. Chem. 264:14646–14652.

    Google Scholar 

  22. Laemmli, U. K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.Nature 227:680–685.

    PubMed  Google Scholar 

  23. Lowry, O. H., N. J. Rosebrough, A. L. Farr, andR. J. Randall. 1951. Protein measurement with the Folin phenol reagent.J. Biol. Chem. 193:265–274.

    PubMed  Google Scholar 

  24. Friberger, P. 1978. Methods for the determination of plasmin, antiplasmin, and plasminogen by means of the substrate S-2251.Haemostasis 7:138–145.

    PubMed  Google Scholar 

  25. Lowry, O. H., N. R. Roberts, M.-L. Wu, W. S. Hixon, andE. J. Crawford. 1953. The quantitative histochemistry of brain; enzyme measurements.J. Biol. Chem. 207:19–37.

    Google Scholar 

  26. Peterkofsky, B., andD. Diegelmann. 1971. Use of a mixture of proteinase-free collagenase for the specific assay of radioactive collagen in the presence of other proteins.Biochemistry 6:988–994.

    Google Scholar 

  27. Chomczynski, P., andN. Sacchi. 1989. Single-step method of RNA isolation by guanidinium thiocyanate-phenol-chloroform extraction.Anal. Biochem. 163:156–162.

    Google Scholar 

  28. Sambrooke, J., E. F. Fritsch, andT. Maniatis. 1989. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor.

    Google Scholar 

  29. Lefebvre, V., C. Peeters-Joris, andG. Vaes. 1990. Modulation by interleukin 1 and tumor necrosis factor α of production of collagenase, tissue inhibitor of metalloproteinase, and collagen types in differentiated and dedifferentiated articular chondrocytes.Biochim. Biophys. Acta 1052:366–378.

    PubMed  Google Scholar 

  30. Guo, J. Z., andH. G. Friesen. 1991. The osteoblastic properties of human osteogenic sarcoma cell line HOS TE85 characterized by morphological, histochemical and molecular biological studies.J. Bone Min. Res. 6:S203 (abstract).

    Google Scholar 

  31. Iamura, K., D. Spriggs, andD. Kufe. 1987. Expression of TNF receptors on human monocytes and internalization of receptor bound ligand.J. Immunol. 139:2989–2992.

    PubMed  Google Scholar 

  32. Heller, R. A., K. Song, N. Fan, andD. J. Chang. 1992. The p70 tumor necrosis factor receptor mediates cytotoxicity.Cell 70:47–56.

    PubMed  Google Scholar 

  33. Hohmann, H.-P., R. Remy, M. Brockhaus, andA. P. G. M. Van Loon. 1989. Two different cell types have different major receptors for human tumor necrosis factor-α.J. Biol. Chem. 264:14927–14934.

    PubMed  Google Scholar 

  34. Vaes, G. 1988. Cellular biology and biochemical mechanisms of bone resorption.Clin. Orthop. Relat. Res. 231:239–272.

    PubMed  Google Scholar 

  35. Thomson, B. M., G. R. Mundy, andT. J. Chambers. 1987. Tumor necrosis factor α and γ induce osteoblastic cells to stimulate osteoclastic bone resorption.J. Immunol 138:775–779.

    PubMed  Google Scholar 

  36. Niedbola, M. J., andM. Stein-Picarella. 1992. Tumor necrosis factor regulation of endothelial cell extracellular proteolysis: The role of urokinase plasminogen activator.Biol. Chem. Hoppe-Seyler. 373:555–566.

    PubMed  Google Scholar 

  37. Marshall, B. C., Q. P. Qu, N. V. Rao, B. R. Brown, andJ. Hoidal. 1992. Pulmonary epithelial cell urokinase-type plasminogen activator. Induction by interleukin-1 and tumor necrosis factor-alpha.J. Biol. Chem. 267:11462–11469.

    PubMed  Google Scholar 

  38. Hamilton, J. A., S. R. Lingelbach, N. C. Partridge, andT. J. Martin. 1984. Stimulation of plasminogen activator in osteoblast-like cells by bone-resorbing hormones.Biochem. Biophys. Res. Commun. 122:230–236.

    PubMed  Google Scholar 

  39. Leloup, G., C. Peeters-Joris, M.-M. Delaisse, G. Opendakker, andG. Vaes. 1991. Tissue and urokinase plasminogen activators in bone tissue and their regulation by parathyroid hormone.J. Bone Min. Res. 6:1081–1089.

    Google Scholar 

  40. Mitchell, P. G., andH. S. Cheang. 1991. Tumor necrosis factor-α and epidermal growth factor regulation of collagenase and stromelysin in adult porcine chondrocytes.J. Cell. Physiol. 149:132–140.

    PubMed  Google Scholar 

  41. Ito, A., T. Sato, T. Iga, andY. Mori. 1990. Tumor necrosis factor bifunctionally regulates matrix metalloproteinases and tissue inhibitor of metalloproteinase (TIMP) production by human fibroblasts.FEBS Lett. 269:93–95.

    PubMed  Google Scholar 

  42. Ahmadzadeh, N., M. Shimgu, andN. Nobunga. 1990. The effect of recombinant tumor necrosis factor-alpha on Superoxide and metalloproteinase production by synovial cells and chondrocytes.Clin. Exp. Rheumatol. 8:387–391.

    PubMed  Google Scholar 

  43. Shimmei, M., K. Masuda, T. Kikuchi, andY. Shimomura. 1989. The role of cytokines in chondrocyte mediated cartilage degradation.J. Rheumatol. (Suppl.) 18:32–34.

    Google Scholar 

  44. Shen, V., G. Kohler, J. J. Jeffrey, andW. A. Peck. 1988. Bone-resorbing agents promote and interferon-β inhibits bone cell collagenase production.J. Bone Min. Res. 3:657–666.

    Google Scholar 

  45. Partridge, N. C., J. J. Jeffrey, L. S. Ehlich, S. L. Teitelbaum, C. Fliszar, H. C. Welgus, andA. J. Kahn. 1987. Hormonal regulation of the production of collagenase and a collagenase inhibitor activity by rat osteogenic sarcoma cells.Endocrinology 120:1956–1961.

    PubMed  Google Scholar 

  46. Lorenzo, J. A., C. C. Pilbeam, J. F. Kalinowski, andM. S. Hibbs. 1992. Production of both 92- and 72-kDa gelatinases by bone cells.Matrix 12:282–290.

    PubMed  Google Scholar 

  47. Nanes, M. S., W. M. McKoy, andS. J. Marx. 1989. Inhibitory effect of tumor necrosis factor-alpha and interferon-gamma on deoxyribonucleic acid and collagen synthesis by rat osteosarcoma cells (ROS 17/2.8).Endocrinology 124:339–345.

    PubMed  Google Scholar 

  48. Scharffetter, K., M. Heckmann, A. Hatamochi, C. Mauch, andB. Stein. 1989. Synergistic effect of tumor necrosis factor-alpha and interferon-gamma on collagen synthesis of human skin fibroblasts in vitro.Exp. Cell Res. 181:409–419.

    PubMed  Google Scholar 

  49. Mauviel, A., J. Heino, V. M. Kahari, D. J. Harthmann, G. Loyau, andJ. P. Pujol. 1991. Comparative effects of interleukin-1 and tumor necrosis factor-alpha on collagen production and corresponding procollagen mRNA levels in human dermal fibroblasts.J. Invest. Dermatol. 96:243–249.

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, UMD-New Jersey Medical School and Graduate School of Biomedical Sciences, 07103-2714, Newark, New Jersey

    Fotinos S. Panagakos & Suriender Kumar

Authors
  1. Fotinos S. Panagakos
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Suriender Kumar
    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

Panagakos, F.S., Kumar, S. Modulation of proteases and their inhibitors in immortal human osteoblast-like cells by tumor necrosis factor-alpha in vitro. Inflammation 18, 243–265 (1994). https://doi.org/10.1007/BF01534267

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01534267

Keywords

Search

Navigation