Skip to main content
SpringerLink
Log in

Cellular approaches to bioreductive drug mechanisms

  • Published:
Cancer and Metastasis Reviews Aims and scope Submit manuscript

Abstract

Mitomycin C is being used as an adjunct to ionizing radiation in the treatment of some solid tumors. A rationale for this is that radioresistant hypoxic cells in solid tumors will have enhanced sensitivity to this bioreductively activated drug, compared to aerobic cells. The role of oxygen concentration and enzymatic drug reduction in bioreductive drug activation have been investigated. Techniques are reviewed for thein vitro determination of the oxygen concentration dependency of bioreductive drug activation. One of these techniques, an open cell suspension system using Chinese hamster ovary cells, is described. Results are shown that indicate that the oxygen concentration dependency of toxicity of mitomycin C and one of its analogues porfiromycin, though qualitatively complementing the oxygen dependency of ionizing radiation toxicity, are not quantitatively optimal. Using a mitomycin C resistant human cell strain (3437T) from a cancer prone family, a possible role for DT-diaphorase, an oxygen insensitive 2-electron transfer enzyme, is suggested. A correlation between a low level of DT-diaphorase in 3437T cells and mitomycin C resistance under aerobic exposure conditions is seen. Under hypoxic exposure conditions this resistance is lost, suggesting 1-electron transfer enzymes control hypoxic cell bioreductive activation. An activation role for DT-diaphorase in mitomycin C toxicity in the treatment of solid tumors is contrasted to a potential detoxification role for the enzyme with other xenobiotics in the cancer prone family phenotype.

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. Suit HD, Miralbell R: Potential impact of improvements in radiation therapy on quality of life and survival. Int J Radiat Oncol Biol Phys 16: 891–895, 1989

    Google Scholar 

  2. Bush RS: Malignancies of the Ovary, Uterus and Cervix. Edward Arnold Ltd, 1979

  3. Moulder JE, Rockwell S: Tumour hypoxia: its impact on cancer therapy. Cancer Metastasis Rev 5: 313–341, 1987

    Google Scholar 

  4. Urtasun RC, Chapman JD, Raleigh JA, Franko AJ, Koch CJ: Binding of3H-misonidazole to solid human tumors as a measure of tumor hypoxia. Int J Radiat Oncol Biol Phys 12: 1263–1269, 1987

    Google Scholar 

  5. Dische S: Keynote address: Hypoxic cell sensitizers: clinical developments. Int J Radiat Oncol Biol Phys 16: 1057–1060, 1989

    Google Scholar 

  6. Jain RK: Delivery of novel therapeutic agents in tumors: physiological barriers and strategies. J Nat Cancer Inst 81: 570–576, 1989

    Google Scholar 

  7. Fowler JF, Adams GE, Denekamp J: Radiosensitizers of hypoxic cells in solid tumors. Cancer Treat Rev 3: 227–256, 1976

    Google Scholar 

  8. Wardman P: The use of nitroaromatic compounds as hypoxic cell radiosensitizers. Current Topics in Rad Res Quarterly 11: 347–398, 1977

    Google Scholar 

  9. Dische S: Chemical sensitizers for hypoxic cells-a decade of experience in clinical radiotherapy. Radiother Oncol 3: 97–115, 1985

    Google Scholar 

  10. Overgaard J, Hansen HS, Andersen AP, Hjelm-Hansen M, Jorgensen K, Sandberg E, Berthelsen A, Hammer R, Pedersen M: Misonidazole combined with radiotherapy in the treatment of carcinoma of the uterine cervix. Int J Radiat Oncol Biol Phys Suppl 16: 1065–1068, 1989

    Google Scholar 

  11. Overgaard J, Sand Hansen H, Lindeløv B, Overgaard M, Jorgensen K, Rasmusson B: Nimorazole as a hypoxic radiosensitizer in the treatment of supraglottic larynx and pharynx carcinoma. First report from the Danish Head And Neck Cancer Study (DAHANCA) protocol 5-85. Radiotherapy Oncology Suppl 20: 143–149, 1991

    Google Scholar 

  12. Sutherland RM: Selective chemotherapy of noncycling cells in anin vitro tumor model. Cancer Res 34: 3501–3503, 1974

    Google Scholar 

  13. Hall EJ, Roizin-Towle L: Hypoxic sensitizer: radiobiological studies at the cellular level. Radiology 117: 453–457, 1975

    Google Scholar 

  14. Moore BA, Palcic B, Skarsgard LD: Radiosensitizing and toxic effects of the 2-nitroimidazole Ro-07-0582 in hypoxic mammalian cells. Radiat Res 67: 459–473, 1976

    Google Scholar 

  15. Taylor YC, Rauth AM: Differences in the toxicity and metabolism of the 2-nitroimidazole misonidazole (Ro-07-0582) in HeLa and Chinese hamster ovary cells. Cancer Res 38: 2745–2752, 1978

    Google Scholar 

  16. Rauth AM: Pharmacology and toxicology of sensitizers: mechanism studies. Int J Radiat Oncol Biol Phys 10: 1293–1300, 1984

    Google Scholar 

  17. Rockwell S, Kennedy KA: Combination therapy with radiation and mitomycin C: preliminary results with EMT6 tumor cellsin vitro andin vivo. Int J Radiat Biol Phys 5: 1673–1676, 1979

    Google Scholar 

  18. Sartorelli AC: Presidential address: Therapeutic attack of hypoxic cells of solid tumors. Cancer Res 48: 775–778, 1988

    Google Scholar 

  19. Zeman EM, Brown JM, Lemmon MJ, Hirst VK, Lee LL: SR-4233: A new bioreductive agent with high selective toxicity for hypoxic mammalian cells. Int J Radiat Oncol Biol Phys 12: 1239–1242, 1986

    Google Scholar 

  20. Brown JM: Redox activation of benzotriazine N-oxides: mechanisms and potential as anticancer drugs. In: Adams GE, Breccia A, Fielden EM, Wardman P (eds) Selective Activation of Drugs by Redox Processes. Plenum Press, London, 1990, pp137–148

    Google Scholar 

  21. Kennedy KA, Rockwell S, Sartorelli AC: Preferential activation of mitomycin C to cytotoxic metabolites by hypoxic tumor cells. Cancer Res 40: 2356–2360, 1980

    Google Scholar 

  22. Rauth AM, Mohindra JK, Tannock IF: Activity of mitomycin C for aerobic and hypoxic cellsin vitro andin vivo. Cancer Res 43: 4154–4158, 1983

    Google Scholar 

  23. Powis G: Free radical formation by antitumor quinones. Free Rad Biol Med 6: 63–110, 1989

    Google Scholar 

  24. Laderoute K, Wardman P, Rauth AM: Molecular mechanisms for the hypoxia-dependent activation of 3-ami-no-1,2,4-benzotriazine-1,4-dioxide (SR4233). Biochemical Pharmacol 37: 1487–1495, 1988

    Google Scholar 

  25. Boag JW: Oxygen diffusion and oxygen depletion problems in radiobiology. In: Ebert M, Howard A (eds) Current Topics in Radiation Research, V. North-Holland Pub, Amsterdam, 1969, pp141–195

    Google Scholar 

  26. Chapman JD, Sturrock J, Boag JW, Crookall JO: Factors affecting the oxygen tension around cells growing in plastic Petri dishes. Int J Radiat Biol 17: 305–328, 1970

    Google Scholar 

  27. Young SD: Hypoxia and Cancer Metastasis. Ph.D. Thesis, University of Toronto, 1990

  28. Whillans DW, Rauth AM: An experimental and analytical study of oxygen depletion in stirred cell suspensions. Radiat Res 84: 97–114, 1980

    Google Scholar 

  29. Marshall RS, Koch CJ, Rauth AM: Measurement of low levels of oxygen and their effect on respiration in cell suspensions maintained in an open system. Radiat Res 108: 91–101, 1986

    Google Scholar 

  30. Fengler JJP: Respiration-induced oxygen gradients in cultured mammalian cells. M.Sc. Thesis, University of British Columbia, 1988

  31. Jones DP, Aw TY, Shan X: Characteristics of hypoxic cells that enhance their susceptibility to chemical injury. In: Adams GE, Breccia A, Fielden EM, Wardman P (eds) Selective Activation of Drugs by Redox Processes. Plenum Press, London, 1990, pp1–9

    Google Scholar 

  32. Koch CJ: A thin-film culturing technique allowing rapid gasliquid equilibration (6sec.) with no toxicity to mammalian cells. Radiat Res 97: 434–442, 1984

    Google Scholar 

  33. Rauth AM, Marshall RS: Mechanisms of activation of mitomycin C and AZQ in aerobic and hypoxic mammalian cells. In: Adams GE, Breccia A, Fielden EM, Wardman P (eds) Selective Activation of Drugs by Redox Processes. Plenum Press, London, 1990, pp113–123

    Google Scholar 

  34. Marshall RS, Rauth AM: Modification of the cytotoxic activity of mitomycin C by oxygen and ascorbic acid in Chinese hamster ovary cells and a repair deficient mutant. Cancer Res 46: 2709–2713, 1986

    Google Scholar 

  35. Marshall RS, Rauth AM: Oxygen and exposure kinetics as factors influencing the cytotoxicity of porfiromycin, a mitomycin C analogue, in Chinese hamster ovary cells. Cancer Res 48: 5655–5659, 1988

    Google Scholar 

  36. Keyes SR, Rockwell S, Sartorelli AC: Porfiromycin as a bioreductive alkylating agent with selective toxicity to hypoxic EMT6 tumor cellsin vivo andin vitro. Cancer Res 45: 3642–3645, 1985

    Google Scholar 

  37. Rockwell S, Keyes SR. Sartorelli AC: Preclinical studies of porfiromycin as an adjunct to radiotherapy. Radiat Res 116: 100–113, 1988

    Google Scholar 

  38. Rockwell S, Nierenburg M, Irvin CG: Effects of the mode of administration of mitomycin on tumor and marrow response and on the therapeutic ratio. Cancer Treat Rep 71: 927–934, 1987

    Google Scholar 

  39. Rockwell S, Sartorelli AC: Mitomycin C and radiation. In: Hill BT, Bellamy AS (eds) Antitumor Drug Radiation Interactions. CRC Press Inc, Boca Raton Florida, 1990, pp125–139

    Google Scholar 

  40. Taylor YC, Rauth AM: Oxygen tension, cellular respiration, and redox state as variables influencing the cytotoxicity of the radiosensitizer misonidazole. Radiat Res 91: 104–123, 1983

    Google Scholar 

  41. Silva ML: One-electron versus two-electron bioreductive cytotoxic mechanisms for hypoxic selective anticancer drugs. Ph.D. Thesis, University of Toronto, 1992

  42. Paterson MC, Middlestadt MV, Weinfeld M, Mirzayans R, Gentner NE: Human cancer-prone disorders, abnormal carcinogenesis response, and defective DNA metabolism. In: Burns FJ, Upton AC, Silini G (eds) Radiation Carcinogenesis and DNA Alterations. Plenum Press, New York, 1986, pp471–498

    Google Scholar 

  43. Marshall RS, Paterson MC, Rauth AM: Deficient activation by a human cell strain leads to mitomycin C resistance under aerobic but not hypoxic conditions. Br J Cancer 59: 341–346, 1989

    Google Scholar 

  44. Dorr RT, Liddil JD, Trent JM: Mitomycin C resistant L-1210 leukemia cells: association with pleiotropic drug resistance. Biochem Pharmacol 36: 3115–3120, 1987

    Google Scholar 

  45. Hoban PR, Robson CN, Davies SM, Hall AG, Catton AR, Hickson ID, Harris AL: Reduced topoisomerase II and elevated α class glutathione S-transferase expression in a multidrug resistant CHO cell line highly cross-resistant to mitomycin C. Biochem Pharmacol 43: 685–693, 1992

    Google Scholar 

  46. Keyes SR, Fracasso PM, Heimbrook DC, Rockwell S, Sligar SG, Sartorelli AC: Role of NADPH: cytochrome C reductase and DT-diaphorase in the biotransformation of mitomycin C. Cancer Res 44: 5638–5643, 1984

    Google Scholar 

  47. Pristos CA, Sartorelli AC: Generation of reactive oxygen radicals through bioactivation of mitomycin antibiotics. Cancer Res 46: 3528–3532, 1986

    Google Scholar 

  48. Marshall RS, Paterson MC, Rauth AM: Studies on the mechanism of resistance to mitomycin C and porfiromycin in a human cell strain derived from a cancer-prone individual. Biochem Pharmacol 41: 1351–1360, 1990

    Google Scholar 

  49. Jaiswal A, McBride OW, Adesnik M, Nebert DW: Human dioxin-inducible cytosolic NAD(P)H: menadione oxidoreductase cDNA sequence and localization of gene to chromosome 16. J Biol Chem 263: 13572–13578, 1988

    Google Scholar 

  50. Jaiswal AK, Burnett P, Adesnik M, McBride OW: Nucleotide and deduced amino acid sequence of human cDNA (NQO2) corresponding to a second member of the NAD(P) H:quinone oxidoreductase gene family. Extensive polymorphism at the NQO2 gene locus on chromosome 6. Biochemistry 29: 1899–1906, 1990

    Google Scholar 

  51. Jaiswal AK: Human NAD(P)H:quinone oxidoreductase (NQO1) gene structure and induction by dioxin. Biochemistry 30: 10647–10653, 1991

    Google Scholar 

  52. Dulhanty AM, Li M, Whitmore GF: Isolation of Chinese hamster ovary cell mutants deficient in excision repair and mitomycin C bioactivation. Cancer Res 49: 117–122, 1989

    Google Scholar 

  53. Dulhanty AM, Whitmore GF: Chinese hamster ovary cell lines resistant to mitomycin C under aerobic but not hypoxic conditions are deficient in DT-diaphorase. Cancer Res 51: 1860–1865, 1991

    Google Scholar 

  54. Begleiter A, Leith M, McClarty G, Beenken S, Goldenberg GJ, Wright JA: Characterization of L5178Y murine lymphoblasts resistant to quinone antitumor agents. Cancer Res 48: 1727–1735, 1988

    Google Scholar 

  55. Begleiter A, Robotham E, Lacey G, Leith MK: Increased sensitivity of quinone resistant cells to mitomycin C. Cancer Lett 45: 173–176, 1989

    Google Scholar 

  56. Siegel D, Gibson NW, Preusch PC, Ross D: Metabolism of mitomycin C by DT-diaphorase: role in mitomycin C-induced DNA damage and cytotoxicity in human colon carcinoma cells. Cancer Res 50: 7483–7489, 1990

    Google Scholar 

  57. Traver RD, Horikoshi T, Danenberg KD, Stadlbauer THW, Danenberg PV, Ross D, Gibson NW: NAD(P)H:quinone oxidoreductase gene expression in human colon carcinoma cells: characterization of a mutation which modulates DT-diaphorase activity and mitomycin C sensitivity. Cancer Res 52: 797–802, 1992

    Google Scholar 

  58. Ross D: DT-diaphorase in activation and detoxification of quinones. This volume

  59. Pan S: The role of NAD(P)H:quinone oxidoreductase in mitomycin C and porfiromycin resistant human colon cancer HCT 116 cells. Proc Amer Assoc Cancer Res 33: 461, 1992

    Google Scholar 

  60. Robertson N, Stratford IJ, Houlbrook S, Carmichael J, Adams GE: The sensitivity of human tumor cells to quinone bioreductive drugs: What role for DT-diaphorase? Biochem Pharmac 44: 409–412, 1992

    Google Scholar 

  61. O'Dwyer PH, Perez RP, Clayton M, Goodwin AK, Hamilton TC: Increased DT-diaphorase activity and cross-resistance to mitomycin C (MMC) in a series of cisplatin-resistant human ovarian carcinoma cell lines. Proc Amer Assoc Cancer Res 33: 461, 1992

    Google Scholar 

  62. Hoban PR, Walton MI, Robson CN, Godden J, Stratford IJ, Workman P, Harris AL, Hickson ID: Decreased NADPH:cytochrome P-450 reductase activity and impaired drug activation in a mammalian cell line resistant to mitomycin C under aerobic but not hypoxic conditions. Cancer Res 50: 4692–4697, 1990

    Google Scholar 

  63. Gustafson DL, Pristos CA: Bioactivation of mitomycin C by xanthine dehydrogenase from EMT6 mouse mammary carcinoma tumors. J Nat Cancer Inst 84: 1180–1185, 1992

    Google Scholar 

  64. Talalay P, Benson AM: Elevation of quinone reductase activity by anticarcinogenic antioxidants. Adv Enz Regul 20: 287–300, 1982

    Google Scholar 

  65. Sun Y: Free radicals, antioxidant enzymes and carcinogenesis. Free Rad Biol Med 8: 583–599, 1990

    Google Scholar 

  66. Favreau LV, Pickett CB: Transcriptional regulation of the rat NAD(P)H:quinone reductase gene. J Biol Chem 266: 4556–4561, 1991

    Google Scholar 

  67. Marshall RS, Paterson MC, Rauth AM: DT-diaphorase activity and mitomycin C sensitivity in non-transformed cell strains derived from members of a cancer-prone family. Carcinogenesis 12: 1175–1180, 1990

    Google Scholar 

  68. Beyer RE, Segura-Aguilar J, Lind C, Castro VM: DT-diaphorase activity in various cells in culture emphasis on induction in ascites hepatoma cells. Chem Scr 27A: 145–150, 1987

    Google Scholar 

  69. Cresteil T, Jaiswal AK: High levels of expression of the NAD(P)H:quinone oxidoreductase (NQOI) gene in tumor cells compared to normal cells of the same origin. Biochemical Pharmacol 42: 1021–1027, 1991

    Google Scholar 

  70. Malkinson AM, Siegel D, Forrest GL, Grazdar AF, Oie HK, Chan DC, Bunn PA, Mabry M, Dykes DJ, Harrison Jr. SD, Ross D: Elevated DT-diaphorase activity and messenger RNA content in human non-small cell lung carcinoma: relationship to the response of lung tumor xenografts to mitomycin C. Cancer Res 52: 4752–4757, 1992

    Google Scholar 

  71. Bailey SM, Suggett N, Walton MI, Workman P: Structureactivity relationships for DT-diaphorase reduction of hypoxic cell directed agents: indolquinones and diaziridinyl benzoquinones. Int J Radiat Biol Phys 22: 649–653, 1992

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Division of Experimental Therapeutics, Ontario Cancer Institute, 500 Sherbourne Street, M4X 1K9, Toronto, Ontario, Canada

    A. Michael Rauth

  2. Department of Medical Biophysics, University of Toronto, 500 Sherbourne Street, M4X 1K9, Toronto, Ontario, Canada

    A. Michael Rauth, Raymond S. Marshall & Bonnie L. Kuehl

Authors
  1. A. Michael Rauth
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Raymond S. Marshall
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Bonnie L. Kuehl
    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

Rauth, A.M., Marshall, R.S. & Kuehl, B.L. Cellular approaches to bioreductive drug mechanisms. Cancer Metast Rev 12, 153–164 (1993). https://doi.org/10.1007/BF00689807

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00689807

Key words

Search

Navigation