The mechanism of cytogenetic genotoxicity (clastogenicity, induction, cell cycle delay) of 10−3 M glutathione in V79-E cells, as described by Thust and Bach (1985), was studied in detail by using different treatment conditions. It was found that I-cystine is the essential cofactor in the incubation system. Catalase, but not superoxide dismutase, abolished the genotoxic effect, and the iron chelator desferoxamine, as well as the hydroxyl radical scavenger mannitol, diminished the activity. It is suggested that glutathione, in combination with V79-E cells and cystine, forms a hydrogen peroxide-generating system which provokes the adverse effects. Glutathione as well as I-cysteine and 2-mercaptopropionylglycine, which were checked for comparison, show a “paradoxic genotoxicity,” i.e., at 10−2 M the effects return almost to the level of controls. Concentration dependence and other criteria of cytogenetic genotoxicity observed with glutathione show obvious similarities to those of other oxidatively acting agents and reveal striking differences to the cytogenetic effects of “typical” genotoxins.
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Abbreviations
- BUdR:
-
5-bromodeoxyuridine
- EMEM:
-
Eagle's minimum essential medium
- GSH:
-
glutathione
- MPG:
-
2-mercaptopropionylglycine
- SCE:
-
sister chromatid exchange
- SOD:
-
superoxide dismutase
- TPA:
-
12-0-tetradecanoylphorbol-13-acetate
References
BIAGLOW, J.E., ISSELS, R.W., GERWECK, L.E., VARNES, M.E., JACOBSON, B., MITCHELL, J.B. and RISSO, A. (1984). Factors influencing the oxidation of cysteamine and other thiols: Implications for hyperthermic sensitization and radiation protection. Radiation Res. 100:298–312.
BIRNBOIM, H.C. (1986). DNA strand breaks in human leukocytes induced by superoxide anion, hydrogen peroxide and tumor promoters are repaired slowly compared to breaks induced by ionizing radiation. Carcinogenesis 7:1511–1517.
BIRNBOIM, H.C. and KANABUS-KAMINSKA, M. (1985). The production of DNA strand breaks in human leukocytes by superoxide anion may involve a metabolic process. Proc. Natl. Acad. Sci. USA 82:6820–6824.
DZARLIEVA-PETRUSEVSKA, R.T. and FUSENIG, N.E. (1985). Tumor promoter 12–0-tetradecanoylphorbol-13-acetate (TPA)-induced chromosome aberrations in mouse keratinocyte cell lines: a possible genetic mechanism of tumor promotion. Carcinogenesis 6:1447–1456.
EMERIT, I. and CERUTTI, P.A. (1981). Tumor promoter phorbol-l2-myristate-l3-acetate induces chromosomal damage via indirect action. Nature 293:144–146.
EMERIT, I. and CERUTTI, P.A. (1982). Tumor promoter phorbol-l2-myristate-l3-acetate induces a clastogenic factor in human leukocytes. Proc. Natl. Acad. Sci. USA 79:7509–7513.
EMERIT, I., LEVY, A. and CERUTTI, P. (1983). Suppression of tumor promoter phorbolmyristate acetate-induced chromosome breakage by antioxidants and inhibitors of arachidonic acid metabolism. Mutat. Res. 110:327–335.
FUJITA, Y., WAKABAYASHI, K., NAGAO, M. and SUGIMURA, T. (1985). Implication of hydrogen peroxide in the mutagenicity of coffee. Mutat. Res. 144:227–230.
GEBHART, E. (1981). Sister chromatid exchange (SCE) and structural chromosome aberration in mutagenicity testing. Hum. Genet. 58:235–254.
GLATT, H.R., PROTIC-SABLJIC, M. and OESHO, F. (1983). Mutagenicity of glutathione and cysteine in the Ames test Science 220:961–963.
HARMAN, L.S., MOTTLEY, C. and MASON, R.P. (1984). Free radical metabolites of 1-cysteine oxidation. J. Biol. Chem. 259:5606–5611.
SSELS, R.D., BOURIER, S., BIAGLOW, J.E., GERWECK, L.E. and WILMANNS, W. (1985). Temperature-dependent influence of thiols upon glutathione levels in Chinese hamster ovary cells at cytotoxic concentrations. Cancer Res. 45:6219–6224.
IWATA, L., SHIBUYA, H., OHKAWA, Y. and INUI, N. (1984). Chromosomal aberrations in V79 cells induced by superoxide radical generated by the hypoxanthine-xanthine oxidase system. Toxicol. Lett. 22:75–81.
LATT, S.A. (1981). Sister chromatid exchange formation. Ann. Rev. Genet. 15:11–55.
MACRAE, W.D. and STICH, H.F. (1979). Induction of sister-chromatid exchanges in Chinese hamster ovary cells by thiol and hydrazine compounds. Mutat. Res. 68:351–365.
MAYR, A., BACHMANN, P.A., BIBRACKI, B. and WITTMANN, G. (1974). Virologische Arbeitsmethoden. Vol. 1, pp. 314–315. VEB Gustav Fischer Verlag, Jena.
NAGASAWA, H. and LITTLE, J.B. (1981). Factors influencing the induction of sister chromatid exchanges in mammalian cells by 12–0-tetradecanoyl-phorbol-13-acetate. Carcinogenesis 2:601–607.
NICOTERA, T.M., BLOCK, A.W., GIBAS, Z. and SANDBERG, A.A. (1985). Induction of superoxide dismutase, chromosomal aberrations and sister-chromatid exchanges by paraquat in Chinese hamster fibroblasts. Mutat. Res. 151:263–268.
PERRY, P.E. (1980). Chemical mutagens and sister-chromatid exchange. In: Chemical Mutagens (deSerres, F.J. and A. Hollaender, eds.) Vol. 6, pp. 1–39.
ROSS, D., MOLDEUS, P., SIES, H. and SMYTH, M.T. (1986). Mechanism and relevance of glutathione mutagenicity. Mutat. Res. 175:127–131.
ROWLEY, D.A. and HALLIWELL, B. (1982). Superoxide-dependent formation of hydroxyl radicals in the presence of thiol compounds. FEBS Lett. 138:33–36.
SANFORD, K.K., PARSHAD, R. and GANTI, R. (1986). Response of human cells in culture to hydrogen peroxide and related free radicals generated by visible light: Relationship to cancer susceptibility. In: Free Radicals, Aging and Degenerative Diseases, pp. 373–394, Alan R. Liss, New York.
SHIBUYA, H., IWATA, K., OHKAWA, Y. and INUI, N. (1985). Histidine increases the frequency of chromosomal aberrations induced by the xanthine oxidase-hypoxanthine system in V79 cells. Toxicol. Lett. 28:117–123.
SOFUNI, T. and ISHIDATE, M. (1984). Induction of chromosomal aberrations in cultured Chinese hamster cells in a superoxide-generating system. Mutat. Res. 140:27–31.
SPEIT, G. (1986). The relationship between the induction of SCEs and mutations in Chinese hamster cells. I. Experiments with hydrogen peroxide and caffeine. Mutat. Res. 174:21–26.
SPEIT, G. and VOGEL, W. (1982). The effect of sulfhydryl compounds on sister-chromatid exchanges. II. The question of cell specificity and the role of H2O2. Mutat. Res. 93:175–183.
STICH, H.F., WEI, L. and LAM, P. (1978). The need for a mammalian test system for mutagens: action of some reducing agents. Cancer Lett. 5:199–204.
TAKAGI, S., SHIKITA, M., TERASIMA, T. and AKABOSHI, S. (1974). Specificity of radioprotective and cytotoxic effects of cysteamine in HeLaS3 cells: generation of peroxide as the mechanism of paradoxical toxicity. Radiation Res. 60:292–301.
TAKAYAMA, S. and SAKANISHI, S. (1977). Differential Giemsa staining of sister chromatids after extraction with acids. Chromosoma 64:109–116.
THUST, R. (1979). Cytogenetic properties of Chinese hamster V79-E cells: G-banding, C-banding, nucleolar organizer regions and sister chromatid exchanges. Exp. Pathol. 17:95–99.
THUST, R. and BACH, B. (1985). Exogenous glutathione induces sister chromatid exchanges, clastogenicity and endoreduplications in V79-E Chinese hamster cells. Cell Biol. Toxicol. 1:123–131.
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Thust, R. The mechanism of cytogenetic genotoxicity of exogenous glutathione in V-79 cells in vitro—implication of hydrogen peroxide and general traits of oxidative chromosome damage. Cell Biol Toxicol 4, 241–257 (1988). https://doi.org/10.1007/BF00119249
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DOI: https://doi.org/10.1007/BF00119249