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Differential protective effects of varying degrees of hypoxia on the cytotoxicities of etoposide and bleomycin

  • Original Articles
  • Bleomycin, Etoposide, Hypoxia
  • Published:
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Summary

Oxygen is thought to be involved both directly and indirectly in the mechanisms of action of several anticancer agents. We studied the effects of various oxygen concentrations on the cytotoxicities of the following drugs: bleomycin (BLM), etoposide (VP-16), doxorubicin (DOX), and mitomycin C (MMC). Human sarcoma cells, MESSA, were exposed to drug for 1 h at one of several oxygen concentrations: less than 1%, 2.5%, 5%, 21%, and 95%. Cytotoxicity was assessed by cellular incorporation of 3H-thymidine into DNA 5 days after drug exposure. Control experiments varying oxygen concentration without drugs demonstrated toxicity only at the highest concentration (95%). Three different responses of drug sensitivity to varying oxygen tensions were observed. BLM, which has been shown to utilize oxygen as a substrate in generating free radicals and producing DNA scission, demonstrated a progressive increase in cytotoxicity over the entire range of increasing oxygen concentrations. This is consistent with the model of a BLM-cation-oxygen complex and catalytic reduction of oxygen. VP-16, which also produces DNA strand breakage but by interaction with topoisomerase II, exhibited a threshold response. VP-16 toxicity was ameliorated by anoxic conditions (less than 1% O2), but not by oxygen concentrations of 2.5%–95%. The reason for this protective effect of anoxia with VP-16 is not clear. In contrast, acute anoxia had no effect on the cytotoxicities of DOX and MMC. We conclude that acute hypoxia protects cells from both BLM and VP-16 but that the nature of that protection is different. VP-16 toxicitiy is blunted only by severe anoxia, wheaeas BLM exhibits a dose response effect over the entire range of oxygen concentrations.

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References

  1. Albertini JP, Garnier-Suillerot A, Tosi L (1982) Iron-bleomycin-DNA-system. Evidence of a long-lived bleomycin-ironoxygen intermediate. Biochem Biophys Res Commun 104: 557–563

    Google Scholar 

  2. Bachur NR, Gordon SL, Gee MV (1978) A general mechanism for microsomal activation of quinone anticancer agents to free radicals. Cancer Res 38: 1745–1750

    Google Scholar 

  3. Bedford JS, Mitchell JB (1974) The effect of hypoxia on the growth and radiation response of mammalian cells in culture. Br J Radiol 47: 687–696

    Google Scholar 

  4. Born R, Hug O, Trott HJR (1976) The effect of prolonged hypoxia on growth and viability of Chinese hamster cells. Int J Radiat Oncol Biol Phys 1: 687–697

    Google Scholar 

  5. Brown JM, Yu NY, Cory MJ, Bicknell RB, Taylor DL (1978) In vivo evaluation of the radiosensitizing and cytotoxic properties of newly synthesized electronaffinic drugs. Br J Cancer 37: 206–211

    Google Scholar 

  6. Burger RM, Peisach J, Horwitz SB (1981) Mechanism of bleomycin action: in vitro studies. Life Sci 28: 715–727

    Google Scholar 

  7. Caspary WJ, Niziak C, Lanzo DA, Friedman R, Bachur NR (1979) Bleomycin A2: a ferrous oxidase. Mol Pharmacol 16: 256–260

    Google Scholar 

  8. Caspary WJ, Lanzo DA, Niziak C (1982) Effect of deoxyribonucleic acid on the production of reduced oxygen by bleomycin and iron. Biochemistry 21: 334–338

    Google Scholar 

  9. Chen GL, Yang L, Rowe TC, Halligan BD, Tewey KM, Liu LF (1984) Nonintercalative antitumor drugs interfere with the breakage-reunion reaction of mammalian DNA topoisomerase II. J Biol Chem 259: 13560–13566

    Google Scholar 

  10. Chien M, Grollman AP, Horwitz SB (1977) Bleomycin-DNA interactions: fluorescence and proton magnetic resonance studies. Biochemistry 16: 3641–3647

    Google Scholar 

  11. Ekimoto H, Kuramochi H, Takahashi K, Matsuda A, Umezawa H (1980) Kinetics of the reaction of bleomycin-Fe(II)-O2 complex with DNA. J Antibiot 33: 426–434

    Google Scholar 

  12. Grollman AP, Takeshita M, Pillai KMR, Hohnson F (1985) Origin and cytotoxic properties of base propenals derived from DNA. Cancer Res 45: 1127–1131

    Google Scholar 

  13. Harker WG, MacKintosh FR, Sikic BI (1983) Development and characterization of a human sarcoma cell line, MES-SA, sensitive to multiple drugs. Cancer Res 43: 4943–4950

    Google Scholar 

  14. Harris JW, Shrieve DC (1979) Effects of Adriamycin and X-rays on euoxic and hypoxic EMT-6 cells in vitro. Int J Radiat Oncol Biol Phys 5: 1245–1248

    Google Scholar 

  15. Hill RP, Bush RS (1977) A new method of determining the fraction of hypoxic cells in a transplantable murine sarcoma. Radiat Res 70: 141–153

    Google Scholar 

  16. Ishida R, Takahashi T (1975) Increased DNA chain breakage by combined action of bleomycin and superoxide radical. Biochem Biophys Res Commun 66: 1432–1438

    Google Scholar 

  17. Kalwinsky DK, Look AT, Ducore J, Fridland A (1983) Effects of the epipodophyllotoxin VP-16-213 on cell cycle traverse, DNA synthesis, and DNA strand size in cultures of human leukemic lymphoblasts. Cancer Res 43: 1592–1597

    Google Scholar 

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

    Google Scholar 

  19. Koch CJ, Druuv J, Frey HE, Snyder RA (1973) Plateau-phase in growth induced by hypoxia. Int J Radiat Biol 23: 67–74

    Google Scholar 

  20. Kuramochi H, Takahashi K, Takita T, Umezawa H (1981) An active intermediate formed in the reaction of bleomycin-Fe(II) complex with oxygen. J Antibiot (Tokyo), 34: 576–582

    Google Scholar 

  21. Loike JD (1984) Podophyllotoxin VP-16-213: a review of their divergent mechanisms of action. Trends Pharmacol Sci 5: 30–33

    Google Scholar 

  22. Loike JD, Horwitz SB (1976) Effect of VP-16-213 on the intracellular degradation of DNA in HeLA cells. Biochemistry 15: 5443–5448

    Google Scholar 

  23. Lown JW, Sim SK (1977) The mechanism of the bleomycininduced cleavage of DNA. Biochem Biophys Res Commun 77: 1150–1157

    Google Scholar 

  24. O'Dwyer PJ, Leyland-Jones B, Alonso MT, Marsoni S, Wittes RE (1985) Drug therapy. Etoposide (VP-16-213): current status of an active anticancer drug. New Engl J Med 312: 692–700

    Google Scholar 

  25. Oppenheimer NJ, Rodriguez LO, Hecht SM (1979) Structural studies of “active complex” of bleomycin: assignment of ligands to the ferrous ion in a ferrous-bleomycin-carbon monoxide complex. Proc Natl Acad Sci USA 76: 5616–5620

    Google Scholar 

  26. Peisach J, Burger RM, Horwitz SB (1982) Oxygen activation and DNA cleavage by iron bleomycin. In: Oxygenases and oxygen metabolism. Academic, New York

    Google Scholar 

  27. Potmesil M, Kirschenbaum S, Israel M, Levin M, Khetarpal VK, Silber R: Relationship of adriamycin concentrations to the DNA lesions induced in hypoxic and euoxic L 1210 cells. Cancer Res 43: 3528–3533

  28. Rauth AM, Mohindra JK, Tannock IF (1983) Activity of mitomycin C for aerobic and hypoxic cells in vitro and in vivo. Cancer Res 43: 4154–4158

    Google Scholar 

  29. Rodriguez LO, Hecht SM (1982) Iron(II)-bleomycin. Biochemical and spectral properties in the presence of radical scavengers. Biochem Biophys Res Comm 104: 1470–1476

    Google Scholar 

  30. Roizin-Towle L, Hall EJ (1978) Studies with bleomycin and misonidazole on aerated and hypoxic cells. Br J Cancer 37: 254–260

    Google Scholar 

  31. Roizin-Towle L, Hall EJ (1979) The effect of bleomycin on aerated and hypoxic cells in vitro, in combination with irradiation. Int J Radiat Oncol Biol Phys 5: 1491–1494

    Google Scholar 

  32. Ross W, Rowe T, Glisson B, Yalowich J, Liu L (1984) Role of topoisomerase II in mediating epipodophyllotoxin-induced DNA cleavage. Cancer Res 44: 5857–5860

    Google Scholar 

  33. Sausville EA, Peisach J, Horwitz SB (1976) A role for ferrous ion and oxygen in the degradation of DNA bleomycin. Biochem Biophys Res Commun 73: 814–822

    Google Scholar 

  34. Sausville EA, Stein RW, Peisach J, Horwitz SB (1978) Properties and products of the degradation of DNA by bleomycin and iron (II). Biochem 17: 2746–2754

    Google Scholar 

  35. Shrieve DC, Deen DF, Harris JEW (1983) Effects of extreme hypoxia on the growth and viability of EMT6/SF mouse tumor cells in vitro. Cancer Res 43: 3521–3527

    Google Scholar 

  36. Sikic BI, Rozencweig M, Carter SK (1985) Bleomycin chemotherapy, Academic, New York

    Google Scholar 

  37. Smith E, Stratford IJ, Adams GE (1979) The resistance of hypoxic mammalian cells to chemotherapeutic agents. Br J Cancer 40: 316

    Google Scholar 

  38. Takeshita M, Grollman AP, Ohtsubo E, Ohtsubo H (1978) Interaction of bleomycin with DNA. Proc Natl Acad Sci USA 75: 5983–5987

    Google Scholar 

  39. Tannock I (1982) Response of aerobic and hypoxic cells in a solid tumor to adriamycin and cyclophosphamide and interaction of the drugs with radiation. Cancer Res 42: 4921–4926

    Google Scholar 

  40. Teicher BA, Lazo JS, Sartorelli AC (1981) Classification of antineoplastic agents by their selective toxicities toward oxygenated and hypoxic tumor cells. Cancer Res 41: 73–81

    Google Scholar 

  41. thomlinson RH, Gray LH (1955) The histological structure of some human lung cancers and the possible implications for radiotherapy. Br J Cancer 9: 539–549

    Google Scholar 

  42. Wozniak AJ, Ross WE (1983) DNA damage as a basis for 4′-dimethylepipodophyllotoxin-9-(4,6-0-ethylidene-β-glucopyranoside) (etoposide) cytotoxicity. Cancer Res 43: 120–124

    Google Scholar 

  43. Zuckerman JE, Raffin TA, Brown JM, Newman RA, Etiz BB, Sikic BI (1986) In vitro selection and characterization of a bleomycin-resistant subline of B16 melanoma. Cancer Res 46: 1748–1752

    Google Scholar 

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Authors and Affiliations

  1. Oncology Division Department of Medicine, Stanford University Medical Center, Room M-211, 94305, Stanford, CA, USA

    Toshitada Yamauchi, Thomas A. Raffin, Phillip Yang & Branimir I. Sikic

Authors
  1. Toshitada Yamauchi
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  2. Thomas A. Raffin
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  3. Phillip Yang
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  4. Branimir I. Sikic
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Additional information

Supported by NIH grant CA-27478 from the US Department of Health and Human Services, and by the American Lung Association. Dr. Sikic is a recipient of a Faculty Development Award in Clinical Pharmacology from the Pharmaceutical Manufacturer's Association Foundation.

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Yamauchi, T., Raffin, T.A., Yang, P. et al. Differential protective effects of varying degrees of hypoxia on the cytotoxicities of etoposide and bleomycin. Cancer Chemother. Pharmacol. 19, 282–286 (1987). https://doi.org/10.1007/BF00261473

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  • DOI: https://doi.org/10.1007/BF00261473

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