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. 1990 Jul;62(1):72–77. doi: 10.1038/bjc.1990.232

The role of glutathione in resistance to cisplatin in a human small cell lung cancer cell line.

C Meijer 1, N H Mulder 1, G A Hospers 1, D R Uges 1, E G de Vries 1
PMCID: PMC1971741  PMID: 2390486

Abstract

The role of glutathione (GSH) in resistance to cisplatin (CDDP) was studied in a human small cell lung carcinoma cell line (GLC4) and a CDDP-resistant subline (GLC4-CDDP). In addition to studying the steady state of GSH, the kinetics of this defence system were also studied via the monitoring of the GSH status of the cells under continuous pressure of CDDP. GLC4-CDDP maintained its elevated GSH level whereas GLC4 (under pressure of CDDP) quickly synthesised GSH to about twice its initial level, corresponding with 80% of the GSH level of GLC4-CDDP. D,L-buthionine-S,R-sulphoximine (BSO) was used to analyse the role of GSH in resistance to CDDP. Pretreatment with BSO (48 h, 50 microM, GSH not detectable) increased the CDDP-induced cytotoxicity 2.8-fold in GLC4-CDDP and 1.7-fold in GLC4. In GLC4 no changes in the amount of platinum (Pt) bound to DNA could be observed after GSH depletion. Changes in formation of interstrand cross-links or the main Pt-containing intrastrand cross-link in digested DNA, the Pt-GG adduct, were also not observed. In GSH depleted GLC4-CDDP cells, an increase in the amount of Pt bound to DNA and in the Pt-GG adduct was observed. Pretreatment with BSO substantially reduced the repair of Pt bound to DNA in both cell lines. We conclude that an increased GSH level and GSH synthesis capacity were demonstrated in CDDP resistant cells. The observations after BSO treatment suggest two roles for GSH in CDDP resistance, namely that of a cytosolic elimination resulting in less DNA platination and a nuclear effect on the formation and repair of DNA platinum adducts.

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Selected References

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  1. Andrews P. A., Velury S., Mann S. C., Howell S. B. cis-Diamminedichloroplatinum(II) accumulation in sensitive and resistant human ovarian carcinoma cells. Cancer Res. 1988 Jan 1;48(1):68–73. [PubMed] [Google Scholar]
  2. Arrick B. A., Nathan C. F. Glutathione metabolism as a determinant of therapeutic efficacy: a review. Cancer Res. 1984 Oct;44(10):4224–4232. [PubMed] [Google Scholar]
  3. Bedford P., Fichtinger-Schepman A. M., Shellard S. A., Walker M. C., Masters J. R., Hill B. T. Differential repair of platinum-DNA adducts in human bladder and testicular tumor continuous cell lines. Cancer Res. 1988 Jun 1;48(11):3019–3024. [PubMed] [Google Scholar]
  4. Behrens B. C., Hamilton T. C., Masuda H., Grotzinger K. R., Whang-Peng J., Louie K. G., Knutsen T., McKoy W. M., Young R. C., Ozols R. F. Characterization of a cis-diamminedichloroplatinum(II)-resistant human ovarian cancer cell line and its use in evaluation of platinum analogues. Cancer Res. 1987 Jan 15;47(2):414–418. [PubMed] [Google Scholar]
  5. Carmichael J., DeGraff W. G., Gazdar A. F., Minna J. D., Mitchell J. B. Evaluation of a tetrazolium-based semiautomated colorimetric assay: assessment of chemosensitivity testing. Cancer Res. 1987 Feb 15;47(4):936–942. [PubMed] [Google Scholar]
  6. Crook T. R., Souhami R. L., Whyman G. D., McLean A. E. Glutathione depletion as a determinant of sensitivity of human leukemia cells to cyclophosphamide. Cancer Res. 1986 Oct;46(10):5035–5038. [PubMed] [Google Scholar]
  7. Eastman A. Cross-linking of glutathione to DNA by cancer chemotherapeutic platinum coordination complexes. Chem Biol Interact. 1987 Mar;61(3):241–248. doi: 10.1016/0009-2797(87)90004-4. [DOI] [PubMed] [Google Scholar]
  8. Eastman A., Schulte N. Enhanced DNA repair as a mechanism of resistance to cis-diamminedichloroplatinum(II). Biochemistry. 1988 Jun 28;27(13):4730–4734. doi: 10.1021/bi00413a022. [DOI] [PubMed] [Google Scholar]
  9. Edgren M., Révész L. Compartmentalised depletion of glutathione in cells treated with buthionine sulphoximine. Br J Radiol. 1987 Jul;60(715):723–724. doi: 10.1259/0007-1285-60-715-723. [DOI] [PubMed] [Google Scholar]
  10. Evans C. G., Bodell W. J., Tokuda K., Doane-Setzer P., Smith M. T. Glutathione and related enzymes in rat brain tumor cell resistance to 1,3-bis(2-chloroethyl)-1-nitrosourea and nitrogen mustard. Cancer Res. 1987 May 15;47(10):2525–2530. [PubMed] [Google Scholar]
  11. Fichtinger-Schepman A. M., Baan R. A., Luiten-Schuite A., van Dijk M., Lohman P. H. Immunochemical quantitation of adducts induced in DNA by cis-diamminedichloroplatinum (II) and analysis of adduct-related DNA-unwinding. Chem Biol Interact. 1985 Nov;55(3):275–288. doi: 10.1016/s0009-2797(85)80135-6. [DOI] [PubMed] [Google Scholar]
  12. Fichtinger-Schepman A. M., van Oosterom A. T., Lohman P. H., Berends F. cis-Diamminedichloroplatinum(II)-induced DNA adducts in peripheral leukocytes from seven cancer patients: quantitative immunochemical detection of the adduct induction and removal after a single dose of cis-diamminedichloroplatinum(II). Cancer Res. 1987 Jun 1;47(11):3000–3004. [PubMed] [Google Scholar]
  13. Green J. A., Vistica D. T., Young R. C., Hamilton T. C., Rogan A. M., Ozols R. F. Potentiation of melphalan cytotoxicity in human ovarian cancer cell lines by glutathione depletion. Cancer Res. 1984 Nov;44(11):5427–5431. [PubMed] [Google Scholar]
  14. Hamilton T. C., Winker M. A., Louie K. G., Batist G., Behrens B. C., Tsuruo T., Grotzinger K. R., McKoy W. M., Young R. C., Ozols R. F. Augmentation of adriamycin, melphalan, and cisplatin cytotoxicity in drug-resistant and -sensitive human ovarian carcinoma cell lines by buthionine sulfoximine mediated glutathione depletion. Biochem Pharmacol. 1985 Jul 15;34(14):2583–2586. doi: 10.1016/0006-2952(85)90551-9. [DOI] [PubMed] [Google Scholar]
  15. Hansson J., Edgren M., Ehrsson H., Ringborg U., Nilsson B. Effect of D,L-buthionine-S,R-sulfoximine on cytotoxicity and DNA cross-linking induced by bifunctional DNA-reactive cytostatic drugs in human melanoma cells. Cancer Res. 1988 Jan 1;48(1):19–26. [PubMed] [Google Scholar]
  16. Hospers G. A., Mulder N. H., de Jong B., de Ley L., Uges D. R., Fichtinger-Schepman A. M., Scheper R. J., de Vries E. G. Characterization of a human small cell lung carcinoma cell line with acquired resistance to cis-diamminedichloroplatinum(II) in vitro. Cancer Res. 1988 Dec 1;48(23):6803–6807. [PubMed] [Google Scholar]
  17. Hospers G. A., de Vries E. G., Mulder N. H. The formation and removal of cisplatin (CDDP) induced DNA adducts in a CDDP sensitive and resistant human small cell lung carcinoma (HSCLC) cell line. Br J Cancer. 1990 Jan;61(1):79–82. doi: 10.1038/bjc.1990.18. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hromas R. A., Andrews P. A., Murphy M. P., Burns C. P. Glutathione depletion reverses cisplatin resistance in murine L1210 leukemia cells. Cancer Lett. 1987 Jan;34(1):9–13. doi: 10.1016/0304-3835(87)90067-x. [DOI] [PubMed] [Google Scholar]
  19. Kraker A. J., Moore C. W. Accumulation of cis-diamminedichloroplatinum(II) and platinum analogues by platinum-resistant murine leukemia cells in vitro. Cancer Res. 1988 Jan 1;48(1):9–13. [PubMed] [Google Scholar]
  20. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  21. Lai G. M., Ozols R. F., Young R. C., Hamilton T. C. Effect of glutathione on DNA repair in cisplatin-resistant human ovarian cancer cell lines. J Natl Cancer Inst. 1989 Apr 5;81(7):535–539. doi: 10.1093/jnci/81.7.535. [DOI] [PubMed] [Google Scholar]
  22. Micetich K., Zwelling L. A., Kohn K. W. Quenching of DNA:platinum(II) monoadducts as a possible mechanism of resistance to cis-diamminedichloroplatinum(II) in L1210 cells. Cancer Res. 1983 Aug;43(8):3609–3613. [PubMed] [Google Scholar]
  23. Pommier Y., Zwelling L. A., Mattern M. R., Erickson L. C., Kerrigan D., Schwartz R., Kohn K. W. Effects of dimethyl sulfoxide and thiourea upon intercalator-induced DNA single-strand breaks in mouse leukemia (L1210) cells. Cancer Res. 1983 Dec;43(12 Pt 1):5718–5724. [PubMed] [Google Scholar]
  24. Richon V. M., Schulte N., Eastman A. Multiple mechanisms of resistance to cis-diamminedichloroplatinum(II) in murine leukemia L1210 cells. Cancer Res. 1987 Apr 15;47(8):2056–2061. [PubMed] [Google Scholar]
  25. Tan K. B., Mattern M. R., Boyce R. A., Schein P. S. Elevated DNA topoisomerase II activity in nitrogen mustard-resistant human cells. Proc Natl Acad Sci U S A. 1987 Nov;84(21):7668–7671. doi: 10.1073/pnas.84.21.7668. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Teicher B. A., Holden S. A., Kelley M. J., Shea T. C., Cucchi C. A., Rosowsky A., Henner W. D., Frei E., 3rd Characterization of a human squamous carcinoma cell line resistant to cis-diamminedichloroplatinum(II). Cancer Res. 1987 Jan 15;47(2):388–393. [PubMed] [Google Scholar]
  27. Tietze F. Enzymic method for quantitative determination of nanogram amounts of total and oxidized glutathione: applications to mammalian blood and other tissues. Anal Biochem. 1969 Mar;27(3):502–522. doi: 10.1016/0003-2697(69)90064-5. [DOI] [PubMed] [Google Scholar]
  28. Waud W. R. Differential uptake of cis-diamminedichloroplatinum (II) by sensitive and resistant murine L1210 leukemia cells. Cancer Res. 1987 Dec 15;47(24 Pt 1):6549–6555. [PubMed] [Google Scholar]
  29. de Jong S., Zijlstra J. G., Timmer-Bosscha H., Mulder N. H., de Vries E. G. Detection of DNA cross-links in tumor cells with the ethidium bromide fluorescence assay. Int J Cancer. 1986 Apr 15;37(4):557–561. doi: 10.1002/ijc.2910370414. [DOI] [PubMed] [Google Scholar]

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