Flow Cytometry pp 343-354 | Cite as

Cytometry, Antitumour Drugs and DNA Topoisomerases

  • P. J. Smith
Conference paper
Part of the NATO ASI Series book series (volume 67)


Flow cytometry has a significant role to play in the of study cellular factors that determine the cytotoxic potential of anticancer drugs. This brief overview provides specific examples of techniques for the analysis of: drug uptake, nuclear localization of drug molecules and drug-induced cell death. Emphasis has been placed on the study of anticancer drugs recognised as potent DNA topoisomerase II poisons together with examples of how target enzyme expression can be analysed in single cells with respect to cell cycle age. An important advantage of the single cell analytical techniques described is the ability to distinguish population heterogeneity, often a confounding factor in the analysis of human tumour specimens.


Median Fluorescence Intensity Lx104 Cell Antitumour Drug Cell Cycle Dependency Human Tumour Specimen 
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  1. Bontemps J, Houssier C, Fredericq E (1975) Physico-chemical study of the complexes of ‘33258 Hoechst’ with DNA and nucleohistone. Nuc Acids Res 2: 971–984CrossRefGoogle Scholar
  2. 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: 4915–4920PubMedGoogle Scholar
  3. Danks MK, Schmidt CA, Ciitain MC, Suttle DP, Beck WT (1988) Altered catalytic activity of and DNA cleavage by DNA topoisomerase II from human leukaemic cells selected for resistance to VM-26. Biochemistry 27: 8861–8869PubMedCrossRefGoogle Scholar
  4. Dive C, Watson JV, Workman P (1990) Multiparametric analysis of cell membrane permeability by two colour flow cytometry with complementary fluorescent probes. Cytometry 11: 244PubMedCrossRefGoogle Scholar
  5. Drake FH, Hofmann GA, Bartus HF, Mattern MR, Crooke ST, Mirabelli CK (1989) Biochemical and pharmacological properties of p170 and p180 forms of topoisomerase II. Biochemistry 28: 8154–8160PubMedCrossRefGoogle Scholar
  6. Ellwart JW, Dormer P (1990) Vitality measurement using spectrum shift in Hoechst 33342 stained cells. Cytometry 11: 239–243PubMedCrossRefGoogle Scholar
  7. Endicott JA, Ling V (1989) The biochemistry of P-glycoprotein-mediated multidrug resistance. Annu Rev Biochem 58: 137–171PubMedCrossRefGoogle Scholar
  8. Epstein RJ, Smith PJ (1988) Estrogen-induced potentiation of DNA damage and cytotoxicity in human breast cancer cells treated with topoisomerase II interactive antitumour drugs. Cancer Res 48: 297–303PubMedGoogle Scholar
  9. Epstein RJ, Smith PJ, Watson JV, Waters C, Bleehen NM (1989) Oestrogen potentiates topoisomerase-II-mediated cytotoxicity in an activated subpopulation of human breast cancer cells: implications for cytotoxic drug resistant in solid tumours. Int J Cancer 44: 501–505PubMedCrossRefGoogle Scholar
  10. Fox ME, Smith PJ (1990) Long-term inhibition of DNA synthesis and the persistence of trapped topo II complexes in determining the toxicity of the antitumour DNA intercalators mAMS A and mitoxantrone. Cancer Res 50: 5813–5818PubMedGoogle Scholar
  11. Heck MM, Hittelman WN, Earnshaw WC (1988) Differential expression of DNA topoisomerases I and II during the eukaryotic cell cycle. Proc Natl Acad Sci USA 85: 1086–1090PubMedCrossRefGoogle Scholar
  12. Kapusinski 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 Pharmac 30: 231–240CrossRefGoogle Scholar
  13. Liu LF (1989) DNA topoisomerase poisons as anti-tumour drugs. Ann Rev Biochem 58:351–375Google Scholar
  14. Lown JW, Hanstock CC (1985) High field 1H-NMR analysis of the 1:1 intercalation complex of the antitumor agent mitoxantrone and the DNA duplex [d(CpGpCpGp)]2- J Biomol Struct Dynam 2: 1097–1106Google Scholar
  15. 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: 9–16PubMedCrossRefGoogle Scholar
  16. Morgan SA, Watson JV, Twentyman PR, Smith PJ (1989) Flow cytometric analysis of Hoechst 33342 uptake as an indicator of multi-drug resistance in human lung cancer. Br J Cancer 60: 282–287PubMedCrossRefGoogle Scholar
  17. Pommier Y, Kerrigan D, Schwartz RE, Swach JA, McCurdy A (1986) Altered DNA topoisomerase II activity in Chinese hamster cells resistant to topoisomerase II inhibitors. Cancer Res 46: 3075–3081PubMedGoogle Scholar
  18. Preisler HD (1978) Alteration of binding of the suprvital dye Hoechst 33342 to human leukemic cells by adriamycin. Cancer Treat Rep 62: 1393–1396PubMedGoogle Scholar
  19. Roberts RA, Cress AE, Dalton WS (1989) Persistent intracellular binding of mitoxantrone in a human colon carcinoma cell line. Biochem Pharmac 38: 4283–4290CrossRefGoogle Scholar
  20. Shenkenberg TD, Von Hoff DD (1986) Mitoxantrone: a new anti- cancer drug with significant clinical activity. Ann Intern Med 105: 67–81PubMedGoogle Scholar
  21. Smith PJ (1990) DNA topoisomerase dysfunction: a new goal for antitumor chemotherapy. Bio Essays 12: 167–172Google Scholar
  22. Smith PJ, Makinson TA (1989) Cellular consequences of overproduction of DNA topoisomerase II in an ataxia-telangiectasia cell line. Cancer Res 49: 1118–1124PubMedGoogle Scholar
  23. 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 Pharmac 40: 2069–2078CrossRefGoogle Scholar
  24. Smith PJ, Morgan SA and Watson JV (1991) Detection of multidrug resistance and quantification of responses of human tumour cells to cytotoxic agents using flow cytometric spectral shift analysis of Hoechst 33342- DNA fluorescence. Cancer Chemother Pharmac 27: 445–450CrossRefGoogle Scholar
  25. Smith PJ, Nakeff A, Watson JV (1985) Flow cytometric detection of changes in the fluorescent emission spectrum of a vital DNA-specific dye in human tumour cells. Exp Cell Res 159: 37–46PubMedCrossRefGoogle Scholar
  26. Tewey KM, Chen GL, Nelson EM, Liu LF (1984) Intercalate antitumour drugs interfere with the breakage-reunion of mammalian DNA topoisomerase II. J Biol Chem 259: 9182–9187PubMedGoogle Scholar
  27. 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: 417–428PubMedCrossRefGoogle Scholar
  28. Wang JC(1985) DNA topoisomerases. Annu Rev Biochem 54:665–697Google Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1993

Authors and Affiliations

  • P. J. Smith
    • 1
  1. 1.Medical Research Council Clinical Oncology and Radiotherapeutics UnitMRC CentreCambridgeUK

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