Pharmaceutical Research

, Volume 14, Issue 5, pp 625–630 | Cite as

Inhibition of Growth of Dictyostelium discoideum Amoebae by Bisphosphonate Drugs Is Dependent on Cellular Uptake

  • Michael J. Rogers
  • Xiaojuan Xiong
  • Xiaohui Ji
  • Jukka Mönkkönen
  • R. Graham G. Russell
  • Michael P. Williamson
  • Frank H. Ebetino
  • Donald J. Watts


Purpose. The aim of the study was to determine whether bisphosphonates are internalised by Dictyosteliumamoebae and whether cellular uptake is required for their growth-inhibitory effects. Bisphosphonates inhibit growth of amoebae of the slime mould Dictyostelium discoideum, by mechanisms that appear to be similar to those that cause inhibition of osteoclastic bone resorption.

Methods. Cell-free extracts prepared from amoebae that had been incubated with bisphosphonates were analysed by 3lP-n.m.r. spectroscopy or ion-exchange f.p.l.c., to identify the presence of bisphosphonates or bisphosphonate metabolites respectively. The growth-inhibitory effect of bisphosphonates towards Dictyostelium amoebae was also examined under conditions in which pinocytosis was inhibited.

Results. All of the bisphosphonates studied were internalised by Dictyostelium amoebae, probably by fluid-phase pinocytosis, and could be detected in cell-free extracts. Amoebae that were prevented from internalising bisphosphonates by pinocytosis were markedly resistant to the growth-inhibitory effects of these compounds. In addition, bisphosphonates encapsulated within liposomes were more potent growth inhibitors of Dictyostelium owing to enhanced intracellular delivery of bisphosphonates.

Conclusions. All bisphosphonates inhibit Dictyostelium growth by intracellular mechanisms following internalisation of bisphosphonates by fluid-phase pinocytosis. It is therefore likely that bisphosphonates also affect osteoclasts by interacting with intracellular, rather than extracellular, processes.

bisphosphonates Dictyostelium endocytosis growth inhibition 31P-n.m.r. osteoclasts 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    H. Fleisch. Drugs 42:919–942 (1991).PubMedGoogle Scholar
  2. 2.
    H. Fleisch. In P. F. Baker, (ed), Handbook of Experimental Pharmacology, vol. 83, Springer, Berlin/Heidelberg, 1988, pp 441–466.Google Scholar
  3. 3.
    A. D. Geddes, S. M. D'Souza, F. H. Ebetino, and K. J. Ibbotson. Bone Mineral Res. 8:265–306 (1994).Google Scholar
  4. 4.
    H. Fleisch. Osteopor. Int. 2:S15–22 (1993).Google Scholar
  5. 5.
    M. J. Rogers, and R. G. G. Russell. In P. T. Sharpe (ed), The Molecular Biology of Paget's Disease. R. G. Landes Co, Austin, TX, pp 147–177, (1996).Google Scholar
  6. 6.
    G. A. Rodan, and H. A. Fleisch. J. Clin. Invest. 97:2692–2696 (1996).PubMedGoogle Scholar
  7. 7.
    D. E. Hughes, K. R. Wright, H. L. Uy, A. Sasaki, T. Yoneda, G. D. Roodman, G. R. Mundy, and B. F. Boyce. J. Bone Miner. Res. 10:1478–1487 (1995).PubMedGoogle Scholar
  8. 8.
    M. J. Rogers, K. M. Chilton, F. Coxon, J. Lawry, M. O. Smith, S. Suri, and R. G. G. Russell. J. Bone Miner. Res. 11:1481–1492 (1996).Google Scholar
  9. 9.
    J. Mönkkönen, M. Taskinen, S. O. K. Auriola, and A. Urtti. J. Drug Targetting 2:299–308 (1994).Google Scholar
  10. 10.
    D. K. Fast, R. Felix, C. Dowse, W. F. Neuman, and H. Fleisch. Biochem. J. 172:97–107 (1978).PubMedGoogle Scholar
  11. 11.
    G. Klein, J.-B. Martin, and M. Satre. Biochemistry 27:1897–1901 (1988).Google Scholar
  12. 12.
    M. J. Rogers, D. J. Watts, R. G. G. Russell, X. Ji, X. Xiong, G. M. Blackburn, A. V. Bayless, and F. H. Ebetino. J. Bone Miner. Res. 9:1029–1039 (1994).PubMedGoogle Scholar
  13. 13.
    M. J. Rogers, X. Xiong, R. J. Brown, D. J. Watts, R. G. G. Russell, A. V. Bayless, and F. H. Ebetino. Molecular Pharmacology 47:398–402 (1995).PubMedGoogle Scholar
  14. 14.
    M. J. Rogers, X. Ji, R. G. G. Russell, G. M. Blackburn, M. P. Williamson, A. V. Bayless, F. H. Ebetino, and D. J. Watts. Dictyostelium discoideum. Biochem. J. 303:303–311 (1994).Google Scholar
  15. 15.
    M. Rogers, R. G. G. Russell, G. M. Blackburn, M. P. Williamson, and D. J. Watts. Biochem. Biophys. Res. Commun. 189:414–423 (1992).PubMedGoogle Scholar
  16. 16.
    M. Clarke, and S. C. Kayman. Meth. Cell Biol. 28:157–176 (1987).Google Scholar
  17. 17.
    G. Klein, D. A. Cotter, J.-B. Martin, and M. Satre. J. Cell Sci. 94:127–134 (1989).Google Scholar
  18. 18.
    C. Gonzalez, G. Klein, and M. Satre. J. Cell Physiol. 144:408–415 (1990).PubMedGoogle Scholar
  19. 19.
    R. R. Sussman, and M. Sussman. Biochem. Biophys. Res. Commun. 29:53–55 (1967).PubMedGoogle Scholar
  20. 20.
    J. Mönkkönen, and T. D. Heath. Calcif. Tiss. Int. 53:139–146 (1993).Google Scholar
  21. 21.
    M. Satre, and J.-B. Martin. Biochem. Biophys. Res. Commun. 132:140–146 (1985).PubMedGoogle Scholar
  22. 22.
    M. H. Chestnut, M. J. Rogers, D. J. Watts, X. Xiong, R. G. G. Russell, F. H. Ebetino, T. L. Grosik, J. L. Finch, J. S. Amburgey, and K. J. Ibbotson. Bone 17(suppl):599 (1995).Google Scholar
  23. 23.
    M. Bof, F. Brenot, C. Gonzalez, G. Klein, J.-B. Martin, and M. Satre. J. Cell Sci. 101:139–144 (1992).Google Scholar
  24. 24.
    F. Brénot, and M. Satre. FEMS Micro. Letts. 109:7–12 (1993).Google Scholar
  25. 25.
    R. Felix, H. L. Guenther, and H. Fleisch. Calcif. Tiss. Int. 36:108–113 (1984).Google Scholar
  26. 26.
    M. Sato, W. Grasser, N. Endo, R. Akins, H. Simmons, D. D. Thompson, E. Golub, and G. A. Rodan. J. Clin. Invest. 88:2095–2105 (1991).PubMedGoogle Scholar
  27. 27.
    A. Carano, S. A. Teitelbaum, J. D. Konsek, P. H. Schlesinger, and H. C. Blair. J. Clin. Invest. 85:456–461 (1990).PubMedGoogle Scholar
  28. 28.
    X. Emonds-Alt, J.-C. Brelière, and R. Roncucci. Biochem. Pharmacol. 34:4043–4049 (1985).CrossRefPubMedGoogle Scholar
  29. 29.
    M. G. Cecchini, and H. Fleisch. J. Bone Miner. Res. 5:1019–1027 (1990).PubMedGoogle Scholar
  30. 30.
    A. M. Flanagan, and T. J. Chambers Calcif. Tiss. Int. 49:407–415 (1991).Google Scholar

Copyright information

© Plenum Publishing Corporation 1997

Authors and Affiliations

  • Michael J. Rogers
    • 1
  • Xiaojuan Xiong
    • 2
  • Xiaohui Ji
    • 2
  • Jukka Mönkkönen
    • 3
  • R. Graham G. Russell
    • 4
  • Michael P. Williamson
    • 2
  • Frank H. Ebetino
    • 5
  • Donald J. Watts
    • 2
  1. 1.Institute for Bone and Joint Medicine, Department of Human Metabolism and Clinical BiochemistryUniversity of Sheffield Medical SchoolUK
  2. 2.Department of Molecular Biology and Biotechnology, University of SheffieldKrebs InstituteSheffieldUK
  3. 3.Department of PharmaceuticsUniversity of KuopioFinland
  4. 4.Institute for Bone and Joint Medicine, Department of Human Metabolism and Clinical BiochemistryUniversity of Sheffield Medical SchoolUK
  5. 5.Procter & Gamble PharmaceuticalsCincinnati

Personalised recommendations