Breast Cancer Research and Treatment

, Volume 82, Issue 3, pp 199–206 | Cite as

Ineffectiveness of Doxorubicin Treatment on Solitary Dormant Mammary Carcinoma Cells or Late-developing Metastases

  • George N. Naumov
  • Jason L. Townson
  • Ian C. MacDonald
  • Sylvia M. Wilson
  • Vivien H.C. Bramwell
  • Alan C. Groom
  • Ann F. Chambers


Breast cancer is noted for long periods of tumor dormancy and metastases can occur many years after treatment. Adjuvant chemotherapy is used to prevent metastatic recurrence but is not always successful. As a model for studying mechanisms of dormancy, we have used two murine mammary carcinoma cell lines: D2.0R/R cells, which are poorly metastatic but form metastases in some mice after long latency times, and D2A1/R cells, which form more numerous metastases much earlier. Previously we identified a surprisingly large population of dormant but viable solitary cells, which persisted in an undivided state for up to 11 weeks after injection of D2.0R/R cells. Dormant cells were also detected for D2A1/R cells, in a background of growing metastases. Here we used this model to test the hypothesis that dormant tumor cells would not be killed by cytotoxic chemotherapy that targets actively dividing cells, and that the late development of metastases from D2.0R/R cells would not be inhibited by chemotherapy that effectively inhibited D2A1/R metastases. We injected mice with D2A1/R or D2.0R/R cells via a mesenteric vein to target liver. We developed a doxorubicin (DXR) treatment protocol that effectively reduced the metastatic tumor burden from D2A1/R cells at 3 weeks. However, this treatment did not reduce the numbers of solitary dormant cells in mice injected with either D2A1/R or D2.0R/R cells. Furthermore, DXR did not reduce the metastatic tumor burden after an 11-week latency period in mice injected with D2.0R/R cells. Thus, apparently effective chemotherapy may spare non-dividing cancer cells, and these cells may give rise to metastases at a later date. This study has important clinical implications for patients being treated with cytotoxic chemotherapy.

breast cancer chemotherapy metastasis murine model tumor dormancy 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Early Breast Cancer Trialists' Collaborative Group: Polychemotherapy for early breast cancer: an overview of the randomized trials. Lancet 352: 930-942, 1998Google Scholar
  2. 2.
    Miller KD, Sledge Jr GW: The role of chemotherapy for metastatic breast cancer. Hematol Oncol Clin North Am 13: 415-434, 1999Google Scholar
  3. 3.
    Tannock IF: Tumor physiology and drug resistance. Cancer Metastasis Rev 20: 123-132, 2001Google Scholar
  4. 4.
    Valeriote F, van Putten L: Proliferation-dependent cytotoxicity of anticancer agents: a review. Cancer Res 35: 2619-2630, 1975Google Scholar
  5. 5.
    Muller I, Niethammer D, Bruchelt G: Anthracycline-derived chemotherapeutics in apoptosis and free radical cytotoxicity. Int J Mol Med 1: 491-494, 1998Google Scholar
  6. 6.
    Gilewski TA, Dang C, Surbone A, Norton L: Cytokinetics. In: Bast RC, Kufe DW, Pollock RE, Weichselbaum RR, Holland JF, Frei E, Gansler TC (eds) Holland and Frei Cancer Medicine. 5th edn, Chapter 38. BC Decker Publishers, Hamilton, Ontario, 2000, pp 511-538Google Scholar
  7. 7.
    Bakemeier RF, Qazi R: Basic concepts of cancer chemotherapy and principles of medical oncology. In: Rubin P, Williams JP (eds) Clinical Oncology. Chapter 8, WB Saunders Company, Philadelphia, 2001, pp 146-159Google Scholar
  8. 8.
    Naumov GN, MacDonald IC, Weinmeister PM, Kerkvliet N, Nadkarni KV, Wilson SM, Morris VL, Groom AC, Chambers AF: Persistence of solitary mammary carcinoma cells in a secondary site: a possible contributor to dormancy. Cancer Res 62: 2162-2168, 2002Google Scholar
  9. 9.
    Morris VL, Koop S, MacDonald IC, Schmidt EE, Grattan M, Percy D, Chambers AF, Groom AC: Mammary carcinoma cell lines of high and low metastatic potential differ not in extravasation but in subsequent migration and growth. Clin Exp Metastasis 12: 357-367, 1994Google Scholar
  10. 10.
    Koop S, MacDonald IC, Luzzi K, Schmidt EE, Kerkvliet N, Morris VL, Grattan M, Khokha R, Chambers AF, Groom AC: Fate of melanoma cells entering the microcirculation: over 80% survive and extravasate. Cancer Res 55: 2520-2523, 1995Google Scholar
  11. 11.
    Luzzi KJ, MacDonald IC, Schmidt EE, Kerkvliet N, Morris VL, Chambers AF, Groom AC: Multistep nature of metastatic inefficiency: dormancy of solitary cells after successful extravasation and limited survival of early micrometastases. Am J Pathol 153: 865-873, 1998Google Scholar
  12. 12.
    Cameron DM, Schmidt EE, Kerkvliet N, Nadkarni KV, Morris VL, Groom AC, Chambers AF, MacDonald IC: Temporal progression of metastasis in lung: cell survival, dormancy, and location dependence of metastatic inefficiency. Cancer Res 60: 2541-2546, 2000Google Scholar
  13. 13.
    Williams SS, Alosco TR, Mayhew E, Lasic DD, Martin FJ, Bankert RB: Arrest of human lung tumor xenograft growth in severe combined immunodeficient mice using doxorubicin encapsulated in sterically stabilized liposomes. Cancer Res 53: 3964-3967, 1993Google Scholar
  14. 14.
    Sugiyama T, Sadzuka Y: Combination of theanine with doxorubicin inhibits hepatic metastasis of M5076 ovarian sarcoma. Clin Cancer Res 5: 413-416, 1999Google Scholar
  15. 15.
    Varghese HJ, Davidson MTM, MacDonald IC, Wilson SM, Nadkarni KV, Groom AC, Chambers AF: Activated Ras regulates the proliferation/apoptosis balance and early survival of developing micrometastases. Cancer Res 62: 887-891, 2002Google Scholar
  16. 16.
    Meltzer A: Dormancy and breast cancer. J Surg Oncol 43: 181-188, 1990Google Scholar
  17. 17.
    Karrison TG, Ferguson DJ, Meier P: Dormancy of mammary carcinoma after mastectomy. J Natl Cancer Inst 91: 80-85, 1999Google Scholar
  18. 18.
    Demicheli R: Tumour dormancy: findings and hypotheses from clinical research on breast cancer. Semin Cancer Biol 11: 297-305, 2001Google Scholar
  19. 19.
    Pocock SJ, Gore SM, Kerr GR: Long term survival analysis: the curability of breast cancer. Stat Med 1: 93-104, 1982Google Scholar
  20. 20.
    Brinkley D, Haybrittle JL: The curability of breast cancer. Lancet 2: 95-97, 1975Google Scholar
  21. 21.
    Duncan W, Kerr GR: The curability of breast cancer. Br Med J 2: 781-783, 1976Google Scholar
  22. 22.
    Brinkley D, Haybittle JL: The curability of breast cancer. World J Surg 1: 287-289, 1977Google Scholar
  23. 23.
    Demicheli R, Abbattista A, Miceli R, Valagussa P, Bonadonna G: Time distribution of the recurrence risk for breast cancer patients undergoing mastectomy: further support about the concept of tumor dormancy. Breast Cancer Res Treat 41: 177-185, 1996Google Scholar
  24. 24.
    Karrison TG, Ferguson DJ, Meier P: Dormancy of mammary carcinoma after mastectomy. J Natl Cancer Inst 91: 80-85, 1999Google Scholar
  25. 25.
    Louwman WJ, Klokman WJ, Coebergh JW: Excess mortality from breast cancer 20 years after diagnosis when life expectancy is normal. Br J Cancer 84: 700-703, 2001Google Scholar
  26. 26.
    Saphner T, Tormey DC, Gray R: Annual hazard rates of recurrence for breast cancer after primary therapy. J Clin Oncol 14: 2738-2746, 1996Google Scholar
  27. 27.
    Gasparini G, Biganzoli E, Bonoldi E, Morabito A, Fanelli M, Boracchi P: Angiogenesis sustains tumor dormancy in patients with breast cancer treated with adjuvant chemotherapy. Breast Cancer Res Treat 65: 71-75, 2001Google Scholar
  28. 28.
    Demicheli R, Retsky MW, Swartzendruber DE, Bonadonna G: Proposal for a new model of breast cancer metastatic Development. Ann Oncol 8: 1075-1080, 1997Google Scholar
  29. 29.
    Retsky MW, Demicheli R, Swartzendruber DE, Bame PD, Wardwell RH, Bonadonna G, Speer JF, Valagussa P: Computer simulation of a breast cancer metastasis model. Breast Cancer Res Treat 45: 193-202, 1997Google Scholar
  30. 30.
    Naumov GN, MacDonald IC, Chambers AF, Groom AC: Solitary cancer cells as a possible source of tumour dormancy? Semin Cancer Biol 11: 271-276, 2001Google Scholar
  31. 31.
    Holmgren L, O'Reilly MS, Folkman J: Dormancy of micrometastases: balanced proliferation and apoptosis in the presence of angiogenesis suppression. Nature Med 1: 149-153, 1995Google Scholar
  32. 32.
    Udagawa T, Fernandez A, Achilles EG, Folkman J, D'Amato RJ: Persistence of microscopic human cancers in mice: alterations in the angiogenic balance accompanies loss of tumor dormancy. FASEB J 16: 1361-1370, 2002Google Scholar
  33. 33.
    Funke I, Schraut W: Meta-analyses of studies on bone marrow micrometastases: an independent prognostic impact remains to be substantiated. J Clin Oncol 16: 557-566, 1998Google Scholar
  34. 34.
    Page DL, Anderson J, Carter BA: Minimal solid tumor involvement of regional and distant sites: when is a metastasis not a metastasis? Cancer 86: 2589-2592, 1999Google Scholar
  35. 35.
    Hermanek P, Hutter RV, Sobin LH, Wittekind C: International Union Against Cancer. Classification of isolated tumor cells and micrometastasis. Cancer 86: 2668-2673, 1999Google Scholar
  36. 36.
    Braun S, Pantel K: Micrometastatic bone marrow involvement: detection and prognostic significance. Med Oncol 16: 154-165, 1999Google Scholar
  37. 37.
    Benson JR, Querci Della Rovere G: Classification of isolated tumor cells and micrometastasis. Cancer 89: 707-709, 2000Google Scholar
  38. 38.
    Muller P, Schlimok G: Bone marrow ‘micrometastases’ of epithelial tumors: detection and clinical relevance. J Cancer Res Clin Oncol 126: 607-618, 2000Google Scholar
  39. 39.
    Pantel K, Cote RJ, Fodstad O: Detection and clinical importance of micrometastatic disease. J Natl Cancer Inst 91: 1113-1124, 1999Google Scholar
  40. 40.
    Diel IJ, Cote RJ: Bone marrow and lymph node assessment for minimal residual disease in patients with breast cancer. Cancer Treat Rev 26: 53-65, 2000Google Scholar
  41. 41.
    Izbicki JR, Pantel K, Hosch SB: Micrometastasis in solid epithelial tumors: impact on surgical oncology. Surgery 131: 1-5, 2002Google Scholar
  42. 42.
    Braun S, Kentenich C, Janni W, Hepp F, de Waal J, Willgeroth F, Sommer H, Pantel K: Lack of effect of adjuvant chemotherapy on the elimination of single dormant tumor cells in bone marrow of high-risk breast cancer patients. J Clin Oncol 18: 80-86, 2000Google Scholar
  43. 43.
    Chambers AF, Naumov GN, Vantyghem SA, Tuck AB: Molecular biology of breast cancer metastasis: clinical implications of experimental studies on metastatic inefficiency. Breast Cancer Res 2: 400-407, 2000Google Scholar

Copyright information

© Kluwer Academic Publishers 2003

Authors and Affiliations

  • George N. Naumov
    • 1
  • Jason L. Townson
    • 1
    • 2
  • Ian C. MacDonald
    • 1
    • 3
  • Sylvia M. Wilson
    • 2
  • Vivien H.C. Bramwell
    • 3
    • 2
    • 4
  • Alan C. Groom
    • 1
  • Ann F. Chambers
    • 1
    • 3
    • 2
  1. 1.Department of Medical BiophysicsUniversity of Western OntarioCanada
  2. 2.London Regional Cancer CentreLondonCanada
  3. 3.Department of OncologyUniversity of Western OntarioCanada
  4. 4.Tom Baker Cancer CentreCalgary, AlbertaCanada

Personalised recommendations