Cell Biology and Toxicology

, Volume 3, Issue 4, pp 347–360 | Cite as

Effect of cellular glutathione depletion on cadmium-induced cytotoxicity in human lung carcinoma cells

  • Yu -Jian Kang
  • M. Duane Eenger

The effect of glutathione depletion on cellular toxicity of cadmium was investigated in a subpopulation (T27) of human lung carcinoma A549 cells with coordinately high glutathione levels and Cd++-resistance. Cellular glutathione levels were depleted by exposing the cells to diethyl maleate or buthionine sulfoximine. Depletion was dose-dependent. Exposure of the cells to 0.5 mM diethyl maleate for 4 hours or to 10 mM buthionine sulfoximine for 8 hours eliminated the threshold for Cd++ cytotoxic effect and deccreased the LD50S. Cells that were pretreated with 0.5 mM diethyl maleate or 10 mM buthionine sulfoximine and then exposed to these same concentrations of diethyl maleate or buthionine sulfoximine during the subsequent assay for colony forming efficiency produced no colonies, reflecting an enhanced sensitivity to these agents at low cell density. Diethyl maleate was found to be more cytotoxic than buthionine sulfoximine. Synergistic cytotoxic effects were observed in the response of diethyl maleate pretreated cells exposed to Cd++. Thus the results demostrated that depletion of most cellular glutathione in A549-T27 cells prior to Cd++ exposure sensitizes them to the agent's cytotoxic effects. Glutathione thus may be involved in modulating the early cellular Cd++ cytotoxic response. Comparison of reduced glutathione levels and of Cd++ cytotoxic responses in buthionine sulfoximine-treated A549-T27 cells with those levels in other, untreated normal and tumor-derived cells suggests that the higher level of glutathione in A549-T27 is not the sole determinant of its higher level of Cd++ resistance.

Key words

A549 cells buthionine sulfoximine Cd++ cytotoxicity diethyl maleate glutathione human tumor cells 





diethyl maleate


dimethyl sulfoxide


reduced glutathione




Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. ANDERSON, M.E. (1985). Detremination of glutathione and glutathione disulfide in biological samples. Method Enzymo. 113:548–555.Google Scholar
  2. BRODIE, A.E. and REED, D.J. (1985). Buthionine sulfoximine inhibition of cystine uptake and glutathione biosynthesis in human lung carcinoma cells. Toxicol. Appl. Phramacol. 77:381–387.Google Scholar
  3. BRUNING, J.L. and KINTZ, B.L. (1977). Computational Handbook of Statistics, pp. 116–131. Scott, Foresman and Company, Glenview, Illinois.Google Scholar
  4. DE GRAFF, W.G. and MITCHELL, J.B. (1985). Glutathione dependence of neocarzinostatin cytotoxicity and mutagenicity in Chinese hamster V-79 cells. Cancer Res. 45:4760–4762.Google Scholar
  5. EENGER, M.D., HILDERBRAND, C.E., WALTERS, R.A, SEARGRAVE, J.C., BARHAM, S.S. and HOAGLUND, H.C. (1984). Molecular and somatic cell genetic analysis of metal resistance mechanisms in mammalian cells. In: Molecular and Cellular Approaches to Understanding Mechanisms of Toxicity. (A.H. Tashjian ed.) pp. 38–62. Published by the Harvard School of Public Health.Google Scholar
  6. EENGER, M.D., TESMER, J.G., TRABVIS, G.L. and BARHAM, S.S> (1986a). Clonal variation of cadmium response in human tumor cell lines. Am. J. Physiol. 250 (Cell Physiol. 19):C256-C263.Google Scholar
  7. EENGER, M.D., HILDERBRAND, C.E., SEARGRAVE, J.C. and TOBEY, R.A. (1986b). Cellular resistance to cadmium. In: Handbook of Experimental Pharmacology, Vol. 80 (E.C. Foulkes ed.) pp. 363–396. Published by Springer-Verlag.Google Scholar
  8. FREEMAN, M.L., MALCOLM, A.W. and MEREDITH, J.J. (1985). Role of glutathione in cell survival after hyperthermic treatment of Chinese hamster ovary cells. Cancer Res. 45:6308–6313.Google Scholar
  9. KETTER, B. (1982). The role of nonenzymatic reactions of glutathione in xenobiotic metabolism. Drug Metabolism Rev. 13(1):161–187.Google Scholar
  10. MEISTER, A. (1985). Methods for the selective modification of glutathione metabolism and study of glutathione transport. Methods Enzymol. 113:571–585.Google Scholar
  11. MITCHELL, J.B., RUSSO, A., KINSELLA, T.J. and GLATSTEIN, E. (1983). Glutathione elevation during thermotolerance induction and thermosensitization by glutathione depletion. Cancer Res. 43:987–991.Google Scholar
  12. MITCHELL, J.B., RUSSO, A., BIGALOW, J.E. and McPHERSON, S. (1983). Cellular glutathione depletion by diethyl maleate or buthionine sulfoximine: no effect of glutathione on the depletion on the oxygen enhancement ratio. Radiation Res. 96:422–428.Google Scholar
  13. REED, D.J. (1984). Cellular defense mechanisms against reactive metabolites. In: Bioactivation of Foreign Compounds (M. W. Anders, ed.) Academic Press, New York.Google Scholar
  14. RICHMAN, P.G. and MEISTER, A. (1975). Regulation of γ-glutamyl-cysteine synthesis by nonallosteric feedback inhibition by glutathione. J. Biol. Chem. 250:1422–1426.Google Scholar
  15. ROIZIN-TOWLE, L. (1985). Selective enhancement of hypoxic cell killing by melphalan via thiol depletuion: in vitro studies with hypoxic cell sensitizers and buthionine sulfoximine. J. Natl. Cancer Inst. 74:151–157.Google Scholar
  16. RUSSO, A., MITCHELL, J.B. and McPHERSON, S. (1984). The effects of glutathione depletion on thermotolerance and heat. Stress protein synthesis. Brit. J. Cancer 49:753–758.Google Scholar
  17. RUSSO, A. and MITCHELL, J.B. (1984). Radiation response of Chinese hamster cells after elevation of intracellular glutathione levels. Int. J. Radiat. Oncol. Biol. Phys. 10:1243–1247.Google Scholar
  18. RUSSO, A., DeGRAFF, W., FRIEDMAN, N. and MITCHELL, J.B. (1986). Selective modulation of glutathione levels in human normal versus tumor cells and subsequent differential response to chemotherapeutic drugs. Cancer Res. 46:2845–2848.Google Scholar
  19. SEAGRAVE, J.C., HILDEBRAND, C.E. and ENGER, M.D. (1983). Effects of cadmium on glutathione metabolism in cadmium sensitive and cadmium resistant Chinese hamster cell lines. Toxicol. 29:101–107.Google Scholar
  20. SHRIEVE, D.L., LI, G.C., ASTROMOFF, A. and HARRIS, J. (1986). Cellular glutathione, thermal sensitivity, and thermotolerance in Chinese hamster fibroblasts and their heat-resistant variants. Cancer Res. 46:1684–1685.Google Scholar
  21. ZAR, J.H. (1984). Biostatistical analysis. pp. 306–326. Prentice-Hall, Inc., Englewood Cliffs, New Jersey.Google Scholar

Copyright information

© Princeton Scientific Publishing Co., Inc 1987

Authors and Affiliations

  • Yu -Jian Kang
    • 1
  • M. Duane Eenger
    • 1
  1. 1.Department of ZoologyIowa State UniversityAmesUSA

Personalised recommendations