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. 1990 Jun 1;268(2):309–315. doi: 10.1042/bj2680309

Expression of human glutathione S-transferases in Saccharomyces cerevisiae confers resistance to the anticancer drugs adriamycin and chlorambucil.

S M Black 1, J D Beggs 1, J D Hayes 1, A Bartoszek 1, M Muramatsu 1, M Sakai 1, C R Wolf 1
PMCID: PMC1131433  PMID: 2194447

Abstract

Adaptation and resistance to chemicals in the environment is a critical part of the evolutionary process. As a result, a wide variety of defence systems that protect cells against chemical insult have evolved. Such chemical resistance mechanisms appear to play a central role in determining the sensitivity of human tumours to treatment with chemotherapeutic drugs. The glutathione S-transferases (GST) are important detoxification enzymes whose over-expression has been associated with drug-resistance. In order to evaluate this possibility we have expressed the human Alpha-class and Pi-class GST cDNAs that encode GST B1B1 and GST pi in the yeast Saccharomyces cerevisiae. The expression of GST B1B1 or GST pi resulted in a marked reduction in the cytotoxic effects of chlorambucil, a bifunctional alkylating agent, and an anthracycline, adriamycin. These data provide direct evidence that the over-expression of GST in cells can confer resistance to anticancer drugs.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Adams K. J., Carmichael J., Wolf C. R. Altered mouse bone marrow glutathione and glutathione transferase levels in response to cytotoxins. Cancer Res. 1985 Apr;45(4):1669–1673. [PubMed] [Google Scholar]
  2. Ammerer G. Expression of genes in yeast using the ADCI promoter. Methods Enzymol. 1983;101:192–201. doi: 10.1016/0076-6879(83)01014-9. [DOI] [PubMed] [Google Scholar]
  3. Arca P., Rico M., Braña A. F., Villar C. J., Hardisson C., Suárez J. E. Formation of an adduct between fosfomycin and glutathione: a new mechanism of antibiotic resistance in bacteria. Antimicrob Agents Chemother. 1988 Oct;32(10):1552–1556. doi: 10.1128/aac.32.10.1552. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Batist G., Tulpule A., Sinha B. K., Katki A. G., Myers C. E., Cowan K. H. Overexpression of a novel anionic glutathione transferase in multidrug-resistant human breast cancer cells. J Biol Chem. 1986 Nov 25;261(33):15544–15549. [PubMed] [Google Scholar]
  5. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bradley G., Juranka P. F., Ling V. Mechanism of multidrug resistance. Biochim Biophys Acta. 1988 Aug 3;948(1):87–128. doi: 10.1016/0304-419x(88)90006-6. [DOI] [PubMed] [Google Scholar]
  7. Buller A. L., Clapper M. L., Tew K. D. Glutathione S-transferases in nitrogen mustard-resistant and -sensitive cell lines. Mol Pharmacol. 1987 Jun;31(6):575–578. [PubMed] [Google Scholar]
  8. CALCUTT G., CONNORS T. A. TUMOUR SULPHYDRYL LEVELS AND SENSITIVITY TO THE NITROGEN MUSTARD MEROPHAN. Biochem Pharmacol. 1963 Aug;12:839–845. doi: 10.1016/0006-2952(63)90114-x. [DOI] [PubMed] [Google Scholar]
  9. Carmichael J., Forrester L. M., Lewis A. D., Hayes J. D., Hayes P. C., Wolf C. R. Glutathione S-transferase isoenzymes and glutathione peroxidase activity in normal and tumour samples from human lung. Carcinogenesis. 1988 Sep;9(9):1617–1621. doi: 10.1093/carcin/9.9.1617. [DOI] [PubMed] [Google Scholar]
  10. Carr B. I., Huang T. H., Buzin C. H., Itakura K. Induction of heat shock gene expression without heat shock by hepatocarcinogens and during hepatic regeneration in rat liver. Cancer Res. 1986 Oct;46(10):5106–5111. [PubMed] [Google Scholar]
  11. Chasseaud L. F. The role of glutathione and glutathione S-transferases in the metabolism of chemical carcinogens and other electrophilic agents. Adv Cancer Res. 1979;29:175–274. doi: 10.1016/s0065-230x(08)60848-9. [DOI] [PubMed] [Google Scholar]
  12. Cowan K. H., Batist G., Tulpule A., Sinha B. K., Myers C. E. Similar biochemical changes associated with multidrug resistance in human breast cancer cells and carcinogen-induced resistance to xenobiotics in rats. Proc Natl Acad Sci U S A. 1986 Dec;83(24):9328–9332. doi: 10.1073/pnas.83.24.9328. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Dulik D. M., Fenselau C., Hilton J. Characterization of melphalan-glutathione adducts whose formation is catalyzed by glutathione transferases. Biochem Pharmacol. 1986 Oct 1;35(19):3405–3409. doi: 10.1016/0006-2952(86)90444-2. [DOI] [PubMed] [Google Scholar]
  14. Duppel W., Lebeault J. M., Coon M. J. Properties of a yeast cytochrome P-450-containing enzyme system which catalyzes the hydroxylation of fatty acids, alkanes, and drugs. Eur J Biochem. 1973 Jul 16;36(2):583–592. doi: 10.1111/j.1432-1033.1973.tb02948.x. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Farber E. The biochemistry of preneoplastic liver: a common metabolic pattern in hepatocyte nodules. Can J Biochem Cell Biol. 1984 Jun;62(6):486–494. doi: 10.1139/o84-066. [DOI] [PubMed] [Google Scholar]
  17. HIRONO I. Non-protein sulphydryl group in the original strain and sub-line of the ascites tumour resistant to alkylating reagents. Nature. 1960 Jun 25;186:1059–1060. doi: 10.1038/1861059a0. [DOI] [PubMed] [Google Scholar]
  18. Habig W. H., Pabst M. J., Jakoby W. B. Glutathione S-transferases. The first enzymatic step in mercapturic acid formation. J Biol Chem. 1974 Nov 25;249(22):7130–7139. [PubMed] [Google Scholar]
  19. Hissin P. J., Hilf R. A fluorometric method for determination of oxidized and reduced glutathione in tissues. Anal Biochem. 1976 Jul;74(1):214–226. doi: 10.1016/0003-2697(76)90326-2. [DOI] [PubMed] [Google Scholar]
  20. Howie A. F., Hayes J. D., Beckett G. J. Purification of acidic glutathione S-transferases from human lung, placenta and erythrocyte and the development of a specific radioimmunoassay for their measurement. Clin Chim Acta. 1988 Sep 30;177(1):65–75. doi: 10.1016/0009-8981(88)90308-7. [DOI] [PubMed] [Google Scholar]
  21. Ito H., Fukuda Y., Murata K., Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983 Jan;153(1):163–168. doi: 10.1128/jb.153.1.163-168.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kano T., Sakai M., Muramatsu M. Structure and expression of a human class pi glutathione S-transferase messenger RNA. Cancer Res. 1987 Nov 1;47(21):5626–5630. [PubMed] [Google Scholar]
  23. Kodate C., Fukushi A., Narita T., Kudo H., Soma Y., Sato K. Human placental form of glutathione S-transferase (GST-pi) as a new immunohistochemical marker for human colonic carcinoma. Jpn J Cancer Res. 1986 Mar;77(3):226–229. [PubMed] [Google Scholar]
  24. Lewis A. D., Hickson I. D., Robson C. N., Harris A. L., Hayes J. D., Griffiths S. A., Manson M. M., Hall A. E., Moss J. E., Wolf C. R. Amplification and increased expression of alpha class glutathione S-transferase-encoding genes associated with resistance to nitrogen mustards. Proc Natl Acad Sci U S A. 1988 Nov;85(22):8511–8515. doi: 10.1073/pnas.85.22.8511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Mannervik B., Danielson U. H. Glutathione transferases--structure and catalytic activity. CRC Crit Rev Biochem. 1988;23(3):283–337. doi: 10.3109/10409238809088226. [DOI] [PubMed] [Google Scholar]
  26. McGown A. T., Fox B. W. A proposed mechanism of resistance to cyclophosphamide and phosphoramide mustard in a Yoshida cell line in vitro. Cancer Chemother Pharmacol. 1986;17(3):223–226. doi: 10.1007/BF00256688. [DOI] [PubMed] [Google Scholar]
  27. Moscow J. A., Fairchild C. R., Madden M. J., Ransom D. T., Wieand H. S., O'Brien E. E., Poplack D. G., Cossman J., Myers C. E., Cowan K. H. Expression of anionic glutathione-S-transferase and P-glycoprotein genes in human tissues and tumors. Cancer Res. 1989 Mar 15;49(6):1422–1428. [PubMed] [Google Scholar]
  28. Paglia D. E., Valentine W. N. Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase. J Lab Clin Med. 1967 Jul;70(1):158–169. [PubMed] [Google Scholar]
  29. Robson C. N., Lewis A. D., Wolf C. R., Hayes J. D., Hall A., Proctor S. J., Harris A. L., Hickson I. D. Reduced levels of drug-induced DNA cross-linking in nitrogen mustard-resistant Chinese hamster ovary cells expressing elevated glutathione S-transferase activity. Cancer Res. 1987 Nov 15;47(22):6022–6027. [PubMed] [Google Scholar]
  30. Shea T. C., Kelley S. L., Henner W. D. Identification of an anionic form of glutathione transferase present in many human tumors and human tumor cell lines. Cancer Res. 1988 Feb 1;48(3):527–533. [PubMed] [Google Scholar]
  31. Smith M. T., Evans C. G., Doane-Setzer P., Castro V. M., Tahir M. K., Mannervik B. Denitrosation of 1,3-bis(2-chloroethyl)-1-nitrosourea by class mu glutathione transferases and its role in cellular resistance in rat brain tumor cells. Cancer Res. 1989 May 15;49(10):2621–2625. [PubMed] [Google Scholar]
  32. Stockman P. K., McLellan L. I., Hayes J. D. Characterization of the basic glutathione S-transferase B1 and B2 subunits from human liver. Biochem J. 1987 May 15;244(1):55–61. doi: 10.1042/bj2440055. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Vernet T., Dignard D., Thomas D. Y. A family of yeast expression vectors containing the phage f1 intergenic region. Gene. 1987;52(2-3):225–233. doi: 10.1016/0378-1119(87)90049-7. [DOI] [PubMed] [Google Scholar]
  35. Wang A. L., Tew K. D. Increased glutathione-S-transferase activity in a cell line with acquired resistance to nitrogen mustards. Cancer Treat Rep. 1985 Jun;69(6):677–682. [PubMed] [Google Scholar]
  36. Wolf C. R., Hayward I. P., Lawrie S. S., Buckton K., McIntyre M. A., Adams D. J., Lewis A. D., Scott A. R., Smyth J. F. Cellular heterogeneity and drug resistance in two ovarian adenocarcinoma cell lines derived from a single patient. Int J Cancer. 1987 Jun 15;39(6):695–702. doi: 10.1002/ijc.2910390607. [DOI] [PubMed] [Google Scholar]
  37. Yoshida Y., Kumaoka H., Sato R. Studies on the microsomal electron-transport system of anaerobically grown yeast. I. Intracellular localization and characterization. J Biochem. 1974 Jun;75(6):1201–1210. doi: 10.1093/oxfordjournals.jbchem.a130503. [DOI] [PubMed] [Google Scholar]
  38. di Ilio C., Sacchetta P., del Boccio G., la Rovere G., Federici G. Glutathione peroxidase, glutathione S-transferase and glutathione reductase activities in normal and neoplastic human breast tissue. Cancer Lett. 1985 Oct;29(1):37–42. doi: 10.1016/0304-3835(85)90120-x. [DOI] [PubMed] [Google Scholar]

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