Skip to main content

Advertisement

Log in

Subcellular localisation of the antitumour drug mitoxantrone and the induction of DNA damage in resistant and sensitive human colon carcinoma cells

  • Original Article
  • Published:
Cancer Chemotherapy and Pharmacology Aims and scope Submit manuscript

Abstract

Cellular uptake and subcellular localisation of the antitumour agent mitoxantrone were studied in a human colon-carcinoma cell line and a mitoxantrone-resistant subline showing features consistent with an atypical multidrug-resistance phenotype involving altered topoisomerase II. Flow cytometry indicated a reduced uptake of mitoxantrone in the resistant line. Confocal microscopy indicated that mitoxantrone-associated fluorescence was primarily found within discrete cytoplasmic inclusions and around the periphery of the nucleus, with low levels being observed within the nucleus. The frequency of cytoplasmic inclusions was reduced in mitoxantrone-resistant cells as compared with parental cells. Fluorescence in cytoplasmic inclusions persisted throughout a 24-h post-treatment period in both cell lines. The results suggest that the persistence of mitoxantrone in cells is a determinant for the continuous induction of DNA damage, perhaps through chronic topoisomerase II trapping, and that modified sequestration may contribute to clinically relevant moderate levels of non-classic multidrug resistance.

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

Access this article

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. Arundel CA, Vines CM, Tofilon PJ (1988) Chromatin modifications associated with N-methylamide-induced radiosensitization of clone A cells. Cancer Res 48: 5669

    PubMed  CAS  Google Scholar 

  2. Beck WT, Cirtain MC, Lefko JL (1983) Energy-dependent reduced drug binding as a mechanism of Vinca alkaloid resistance in human leukaemic lymphoblasts. Mol Pharmacol 24: 485

    PubMed  CAS  Google Scholar 

  3. Bell DH (1988) Characterization of the fluorescence of the antitumour agent, mitoxantrone. Biochim Biophys Acta 949: 132

    PubMed  CAS  Google Scholar 

  4. Bowden GT, Roberts R, Alberts DS, Peng Y-M, Garcia D (1985) Comparative molecular pharmacology in leukaemic L1210 cells of the anthracene anticancer drugs mitoxantrone and bisantrene. Cancer Res 45: 4915

    PubMed  CAS  Google Scholar 

  5. Chegini N, Safa AR (1987) Influence of mitoxantrone on nucleolar function in MDA-MB-231 human breast cancer cell line. Cancer Lett 37: 327

    Article  PubMed  CAS  Google Scholar 

  6. Crespi MD, Ivanier SE, Genovese J, Baldi A (1986) Mitoxantrone affects topoisomerase activities in human breast cancer cells. Biochem Biophys Res Commun 136: 521

    Article  PubMed  CAS  Google Scholar 

  7. Cress AE, Roberts RA, Bowden GT, Dalton WS (1988) Modification of keratin by the chemotherapeutic drug mitoxantrone. Biochem Pharmacol 37: 3043

    Article  PubMed  CAS  Google Scholar 

  8. Dalton WS, Cress AE, Alberts DS, Trent JM (1988) Cytogenetic and phenotypic analysis of a human colon carcinoma cell line resistant to mitoxantrone. Cancer Res 48: 1882

    PubMed  CAS  Google Scholar 

  9. Danks MK, Yalowich JC, Beck WT (1987) Atypical multiple drug resistance in a human leukaemic cell line selected for resistance for teniposide (VM-26). Cancer Res 47: 1297

    PubMed  CAS  Google Scholar 

  10. Egorin MJ, Hildebrand RC, Cimino EF, Bachur NR (1974) Cytofluorescence localisation of Adriamycin and daunorubicin. Cancer Res 34: 2243

    PubMed  CAS  Google Scholar 

  11. Endicott JA, Ling V (1989) The biochemistry of P-glycoproteinmediated multidrug resistance. Annu Rev Biochem 58: 137

    Article  PubMed  CAS  Google Scholar 

  12. Epstein RJ, Smith PJ (1988) Estrogen-induced potentiation of DNA damage and cytotoxicity in human breast cancer cells treated with topoisomerase-II interactive intercalative drugs. Cancer Res 48: 297

    PubMed  CAS  Google Scholar 

  13. Estey EH, Keating MJ, McCredie KB, Bodey GP, Freirich EJ (1982) Phase II trial of dihydroxyanthracenedione in acute leukaemia. Proc Am Assoc Cancer Res 23: 113

    Google Scholar 

  14. Fox ME, Smith PJ (1990) Long-term inhibition of DNA synthesis and the persistence of trapped topoisomerase II complexes in determining the toxicity of the antitumor DNA intercalators mAMSA and mitoxantrone. Cancer Res 50: 5813

    PubMed  CAS  Google Scholar 

  15. Gervasoni JE Jr, Fields SZ, Krishna S, Baker MA, Rosado M, Thurasamy K, Hindenburg AA, Taub RN (1991) Subcellular distribution of daunorubicin in P-glycoprotein-positive and -neggative drug-resistant cell lines using laser-assisted confocal microscopy. Cancer Res 51: 4955

    PubMed  CAS  Google Scholar 

  16. Ho AD, Seither E, Ma DDF, Prentice HG (1987) Mitoxantrone-induced toxicity and DNA strand breaks in leukaemic cells. Br J Haematol 65: 51

    Article  PubMed  CAS  Google Scholar 

  17. Kanter PM, Schwartz HS (1982) A fluorescence enhancement assay for cellular DNA damage. Mol Pharmacol 22: 145

    PubMed  CAS  Google Scholar 

  18. Kapuscinski J, Darzynkiewicz Z, Traganos F, Melamed MR (1981) Interaction of a new antitumour agent, l-4-dihydroxy-5,8-bis[(2-(2-hydroxyethyl)amino)-ethyl)amino]-9,10-anthracenedione, with nucleic acids. Biochem Pharmacol 30: 231

    Article  PubMed  CAS  Google Scholar 

  19. Krishan A, Ganapathi R (1980) Laser flow studies on the intracellular fluorescence of anthracyclines. Cancer Res 40: 3895

    PubMed  CAS  Google Scholar 

  20. Liley DTJ, Wiggins PM, Baguley BC (1989) Localization of a nonintercalative DNA binding antitumour drug in mitochondria: relationship to multidrug resistance. Eur J Cancer Clin Oncol 25: 1287

    Article  PubMed  CAS  Google Scholar 

  21. Lin C-W, Shulok JR, Kirley SD, Cincotta L, Foley JW (1991) Lysosomal localisation and mechanism of uptake of Nile blue photosensitizers in tumor cells. Cancer Res 51: 2710

    PubMed  CAS  Google Scholar 

  22. Liu LF (1989) DNA topoisomerase poisons as antitumour drugs. Annu Rev Biochem 58: 351

    Article  PubMed  CAS  Google Scholar 

  23. Lown JW, Hanstock CC, Bradley RD, Scraba DG (1983) Interactions of the antitumor agents mitoxantrone and bisantrene with deoxyribonucleic acids studied by electron microscopy. Mol Pharmacol 25: 178

    Google Scholar 

  24. Minford J, Pommier Y, Filipski J, Kohn KW, Kerrigan D, Mattern M, Michaels S, Schwartz R, Zwelling LA (1986) Isolation of intercalator-dependent protein-linked DNA strand cleavage activity from cell nuclei and identification as topoisomerase II. Biochemistry 25: 9

    Article  PubMed  CAS  Google Scholar 

  25. Noguchi P, Wallace R, Johnson J, Earley EM, O’Brien S, Ferrone S, Pellegrino MA, Milstien J, Needy C, Browne W, Petricciani J (1979) Characterization of WiDr: a human colon carcinoma cell line. In Vitro 15: 401

    Article  CAS  Google Scholar 

  26. Roberts RA, Cress AE, Dalton WS (1989) Persistent intracellular binding of mitoxantrone in a human colon carcinoma cell line. Biochem Pharmacol 38: 4283

    Article  PubMed  CAS  Google Scholar 

  27. Rowe TC, Chen GL, Hsiang Y-H, Liu LF (1986) DNA damage by antitumour acridines mediated by mammalian DNA topoisomerase II. Cancer Res 46: 2021

    PubMed  CAS  Google Scholar 

  28. Smith PJ, Makinson TA (1989) Cellular consequences of over-production of DNA topoisomerase II in an ataxia-telangiectasia cell line. Cancer Res 49: 1118

    PubMed  CAS  Google Scholar 

  29. Smith PJ, Morgan SA, Fox ME, Watson JV (1990) Mitoxantrone-DNA binding and the induction of topoisomerase II associated DNA damage in multi-drug resistant small cell lung cancer cells. Biochem Pharmacol 40: 2069

    Article  PubMed  CAS  Google Scholar 

  30. Smith PJ, Sykes HR, Fox ME, Furlong IJ (1992) Subcellular distribution of the anticancer drug mitoxantrone in human and drug-resistant murine cells analyzed by flow cytometry and confocal microscopy and its relationship to the induction of DNA damage. Cancer Res 52: 1

    Google Scholar 

  31. Smyth JF, Cornbleet MA, Stuart-Harris RC, Smith IE, Coleman RE, Rubens RD, McDonald M, Mouridsen HT, Rainer H, Oosterom AT van (1984) Mitoxantrone as first-line chemotherapy for advanced breast cancer: results of a European collaborative study. Semin Oncol 11: 15

    PubMed  CAS  Google Scholar 

  32. Tewey KM, Chen GL, Nelson EM, Liu LF (1984) Intercalative antitumour drugs interfere with the breakage-reunion of mammalian DNA topoisomerase II. J Biol Chem 259: 9182

    PubMed  CAS  Google Scholar 

  33. Wallace RE, Lindh D, Durr FE (1987) Development of resistance and characteristics of a human colon carcinoma subline resistant to mitoxantrone in vitro. Cancer Invest 5: 417

    Article  PubMed  CAS  Google Scholar 

  34. Watson JV (1978) A linear transform of the multi-target survival curve. Br J Radiol 51: 534

    Article  PubMed  CAS  Google Scholar 

  35. White JG, Amos WB, Fordham M (1987) Evaluation of confocal versus conventional imaging of biological structures by fluorescence light microscopy. J Cell Biol 105: 41

    Article  PubMed  CAS  Google Scholar 

  36. Yanowich S, Taub RN (1983) Differences in daunomycin retention in sensitive and resistant P388 leukemic cells as determined by digitized video fluorescence microscopy. Cancer Res 41: 67

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Fox, M.E., Smith, P.J. Subcellular localisation of the antitumour drug mitoxantrone and the induction of DNA damage in resistant and sensitive human colon carcinoma cells. Cancer Chemother. Pharmacol. 35, 403–410 (1995). https://doi.org/10.1007/s002800050254

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s002800050254

Key words

Navigation