A comparison of the in vitro genotoxicity of anticancer drugs idarubicin and mitoxantrone.
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Idarubicin is an anthracycline antibiotic used in cancer therapy. Mitoxantrone is an anthracycline analog with presumed better antineoplastic activity and lesser toxicity. Using the alkaline comet assay we showed that the drugs at 0.01-10 μM induced DNA damage in normal human lymphocytes. The effect induced by idarubicin was more pronounced than by mitoxantrone (P < 0.001). The cells treated with mitoxantrone at 1 μM were able to repair damage to their DNA within a 30-min incubation, whereas the lymphocytes exposed to idarubicin needed 180 min. Since anthracyclines are known to produce free radicals, we checked whether reactive oxygen species might be involved in the observed DNA damage. Catalase, an enzyme inactivating hydrogen peroxide, decreased the extent of DNA damage induced by idarubicin, but did not affect the extent evoked by mitoxantrone. Lymphocytes exposed to the drugs and treated with endonuclease III or formamidopyrimidine-DNA glycosylase (Fpg), enzymes recognizing and nicking oxidized bases, displayed a higher level of DNA damage than the untreated ones. 3-Methyladenine-DNA glycosylase II (AlkA), an enzyme recognizing and nicking mainly methylated bases in DNA, increased the extent of DNA damage caused by idarubicin, but not that induced by mitoxantrone. Our results indicate that the induction of secondary malignancies should be taken into account as side effects of the two drugs. Direct strand breaks, oxidation and methylation of the DNA bases can underlie the DNA-damaging effect of idarubicin, whereas mitoxantrone can induce strand breaks and modification of the bases, including oxidation. The observed in normal lymphocytes much lesser genotoxicity of mitoxantrone compared to idarubicin should be taken into account in planning chemotherapeutic strategies.
- 1. Gentile, J.M., Rahimi, S., Zwiesler, J., Gentile, G.J. & Ferguson, L.R. (1998) Effects of selected antimutagens on the genotoxicity of antitumor agents. Mutat. Res. 402, 289-298.
- 2. Kellogg, G.E., Scarsdale, J.N. & Fornari, F.A. (1998) Identification and hydropathic characterization of structural features affecting sequence specificity for doxorubicin intercalation into DNA double-stranded polynucleotides. Nucleic Acids Res. 26, 4721-4732.
- 3. Zhang, H.M. & Li, N.Q. (2000) Electrochemical studies of the interaction of adriamycin to DNA. J. Pharm. Biomed. Anal. 22, 67-73.
- 4. Tewey, K.M., Rowe, T.C., Yang, L., Halligan, B.D. & Liu, L.F. (1984) Adriamycin-induced DNA damage mediated by mammalian DNA topoisomerase II. Science. 226, 466-468.
- 5. Schneider, E., Hsiang, Y. & Liu, L.F. (1990) DNA topoisomerases as anticancer drug targets. Adv. Pharmacol. 21, 149-183.
- 6. Bridewell, D.J., Finlay, G.J. & Baguley, B.C. (1997) Differential actions of aclarubicin and doxorubicin: The role of topoisomerase I. Oncol. Res. 9, 535-542.
- 7. Abraham, R., Basser, R.L. & Green, M.D. (1996) A risk-benefit assessment of anthracycline antibiotics in antineoplastic therapy. Drug Safety. 15, 406-429.
- 8. Faure, H., Mousseau, M., Cadet, J., Guimier, C., Tripier, M., Hida, H. & Favier, A. (1998) Urine 8-oxo-7,8-dihydro-2-deoxyguanosine vs. 5-(hydroxymethyl)uracil as DNA oxidation marker in adriamycin-treated patients. Free Radical Res. 28, 377-382.
- 9. Stathopoulos, G.P., Malamos, N.A., Dontas, I., Deliconstantinos, G., Perrea-Kotsarelis, D. & Karayannacos, P.E. (1998) Inhibition of adriamycin cardiotoxicity by 5-fluorouracil: A potential free oxygen radical scavenger. Anticancer Res. 18, 4387-4392.
- 10. Falcone, G., Filippelli, W., Mazzarella, B., Tufano, R., Mastronardi, P., Filippelli, A., Berrino, L. & Rossi, F. (1998) Cardiotoxicity of doxorubicin: Effects of 21-aminosteroids. Life Sci. 63, 1525-1532.
- 11. Samelis, G.F., Stathopoulos, G.P., Kotsarelis, D., Dontas, I., Frangia, C. & Karayannacos, P.E. (1998) Doxorubicin cardiotoxicity and serum lipid increase is prevented by dextrazoxane (ICRF-187). Anticancer Res. 18, 3305-3309.
- 12. Platel, D., Pouna, P., Bonoron-Adele, S. & Robert, J. (1999) Comparative cardiotoxicity of idarubicin and doxorubicin using the isolated perfused rat heart model. Anticancer Drugs 10, 671-676.
- 13. George, J.W., Ghate, S., Matson, S.W. & Besterman, J.M. (1992) Inhibition of DNA helicase II unwinding and ATPase activities by DNA-interacting ligands. Kinetics and specificity. J. Biol. Chem. 267, 10683-10689.
- 14. Linassier, C., Barin, C., Calais, G., Letortorec, S., Bremond, J.L., Delain, M., Petit, A., Georget, M.T., Cartron, G., Raban, N., Benboubker, L., Leloup, R., Binet, C., Lamagnere, J.P. & Colombat, P. (2000) Early secondary acute myelogenous leukemia in breast cancer patients after treatment with mitoxantrone, cyclophosphamide, fluorouracil and radiation therapy. Ann. Oncol. 11, 1289-1294.
- 15. Singh, N.P., McCoy, T., Tice, R.R. & Schneider, E.L. (1988) A simple technique for quantitation of low levels of DNA damage in individual cells. Exp. Cell Res. 175, 184-192.
- 16. Collins, A.R., Duthie, S.J. & Dobson, V.L. (1993) Direct enzymatic detections of endogenous base damage in human lymphocyte DNA. Carcinogenesis. 14, 1733-1735.
- 17. Fisher, G.R. & Patterson, L.H. (1991) DNA strand breakage by peroxidase-activated mitoxantrone. J. Pharm. Pharmacol. 43, 65-68.
- 18. Reszka, K.J., Matuszak, Z. & Chignell, C.F. (1997) Lactoperoxidase-catalyzed oxidation of the anticancer agent mitoxantrone by nitrogen dioxide radicals. Chem. Res. Toxicol. 10, 1325-1330.
- 19. Borchmann, P., Hubel, K., Schnell, R. & Engert, A. (1997) Idarubicin: A brief overview on pharmacology and clinical use. Int. J. Clin. Pharmacol. Ther. 35, 80-83.
- 20. Duthie, S.J. & Collins, A.R. (1997) The influence of cell growth, detoxifying enzymes and DNA repair of human hydrogen peroxide-mediated DNA damage (measured using the comet assay) in human cells. Free Radical Biol. Med. 22, 717-724.
- 21. David-Cordonnier, M.-H., Laval, J. & O’Neill, P. (2000) Clustered DNA damage, influence on damage excision by XRS5 nuclear extracts and Escherichia coli Nth and Fpg proteins. J. Biol. Chem. 275, 11865-11873.
- 22. Boiteux, S., Gajewski, E., Laval, J. & Dizdaroglu, M. (1992) Substrate specificity of the Escherichia coli Fpg protein (formamidopyrimidine-DNA glycosylase): Excision of purine lesions in DNA produced by ionizing radiation or photosensitization. Biochemistry. 31, 106-110.
- 23. Tchou, J., Kasai, H., Shibutani, S., Chung, M.H., Laval, J., Grollman, A.P. & Nishimura, S. (1991) 8-Oxoguanine (8hydroxyguanine) DNA glycosylase and its substrate specificity. Proc. Natl. Acad. Sci. U.S.A. 88, 4690-4694.
- 24. Laval, J. (1977) Two enzymes are required for strand incision in repair of alkylated DNA. Nature 269, 829-832.
- 25. Tudek, B., VanZeeland, A.A., Kusmierek, J.T. & Laval, J. (1998) Activity of Escherichia coli DNA-glycosylases on DNA damaged by methylating and ethylating agents and influence of 3-substituted adenine derivatives. Mutat. Res. 407, 169-176.
- 26. Klaude, M., Eriksson, S., Nygren, J. & Ahnstrom, G. (1996) The comet assay: Mechanisms and technical considerations. Mutat. Res. 363, 89-96.
- 27. Blasiak, J. & Kowalik, J. (2000) A comparison of the in vitro genotoxicity of tri- and hexavalent chromium. Mutat. Res. 469, 135-145.
- 28. Ashby, J.A., Tinwell, H., Lefevre, P.A. & Brown, M.A. (1995) The single cell gel electrophoresis assay for induced DNA damage (comet assay): Measurement of tail length and moment. Mutagenesis. 10, 85-90.
- 29. Hande, K.R. (1998) Clinical applications of anticancer drugs targeted to topoisomerase II. Biochim. Biophys. Acta. 1400, 173-184.
- 30. Shackelford, R.E., Kaufmann, W.K. & Paules, R.S. (2000) Oxidative stress and cell cycle checkpoint function. Free Radical Biol. Med. 28, 1387-1404.
- 31. Dizdaroglu, M., Rao, G., Halliwell, B. & Gajewski, E. (1991) Damage to the DNA bases in mammalian chromatin by hydrogen peroxide in the presence of ferric and cupric ions. Arch. Biochem. Biophys. 285, 317-324.
- 32. Imlay, J.A., Chin, S.M. & Linn, S. (1991) Toxic DNA damage by hydrogen peroxide through the Fenton reaction in vivo and in vitro. Science. 240, 640-642.
- 33. Zhu, B.Z., Kitrossky, N. & Chevion, M. (2000) Evidence for production of hydroxyl radicals by pentachlorophenol metabolites and hydrogen peroxide: A metal-independent organic Fenton reaction. Biochem. Biophys. Res. Commun. 270, 942-946.
- 34. Termini, J. (2000) Hydroperoxide-induced DNA damage and mutations. Mutat. Res. 450, 107-124.
- 35. Capranico, G., Riva, A., Tinelli, S., Dasdia, T. & Zunino, F. (1987) Markedly reduced levels of anthracycline-induced DNA strand breaks in resistant P388 leukemia cells and isolated nuclei. Cancer Res. 47, 3752-3756.
- 36. Senkal, M., Tonn, J.C., Schonmayr, R., Schachenmayr, W., Eickhoff, U., Kemen, M. & Kollig, E. (1997) Mitoxantrone-induced DNA strand breaks in cell-cultures of malignant human astrocytoma and glioblastoma tumors. J. Neurooncol. 32, 203-208.
- 37. Kolodziejczyk, P., Reszka, K. & Lown, J.W. (1988) Enzymatic oxidative activation and transformation of the antitumor agent mitoxantrone. Free Radical Biol. Med. 5, 13-25.
- 38. Novak, R.F. & Kharasch, E.D. (1985) Mitoxantrone: Propensity for free radical formation and lipid peroxidation - implications for cardiotoxicity. Invest. New Drugs. 3, 95-99.
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