Skip to main content
NCBI home page
Journal of Cancer Research and Clinical Oncology logoLink to Journal of Cancer Research and Clinical Oncology
. 1991 Jul;117(4):313–320. doi: 10.1007/BF01630713

Sensitization of human melanoma cells to melphalan cytotoxicity by Adriamycin and carmustine

Vesna Jevtorić-Todorović 1, Thomas M Guenthner 1,✉
PMCID: PMC12201648  PMID: 2066351

Abstract

Exposure of cultured human melanoma cells from three different cell lines to Adriamycin and carmustine at non-cytotoxic (micromolar) concentrations results in a rapid, reversible depletion of cellular glutathione; maximal depletion is achieved within 1 h, and glutathione levels recover within 2–3 h. Glutathione depletion is accompanied by an enhancement of the cytotoxic effects of the alkylating agent melphalan, which ranges from 15-to 55-fold. These results suggest that the combination of Adriamycin and carmustine may provide a rational drug combination for the rapid depletion of glutathione from malignant melanoma, thereby sensitizing these tumor cells to alkylating agent cytotoxicity.

Key words: Adriamycin, Carmustine, Melphalan, Glutathione, Melanoma

Abbreviations

LPAM

L-phenylalanine mustard, melphalan

GSH

reduced glutathione

ADR

Adriamycin

BCNU

N,N-bis-(2-chloroethyl)-N-nitrosourea, carmustine

GSSG

glutathione disulfide

MEL2

the cultured human melanoma cell line UISO-MEL2

LC90

concentration of drug resulting in 90% inhibition of clonogenicity

HPLC

high-pressure liquid chromatography

Footnotes

These studies are part of a thesis submitted by V.T. to the Graduate College of the University of Illinois at Chicago, in partial fulfillment of the requirements for the Ph.D. degree. Supported by PHS grant CA51218 and RCDA award KO4 CAO1287 (to T.M.G.) from the National Cancer Institute

References

  1. Arrick BA, Nathan CF (1984) Glutathione metabolism as a determinant of therapeutic efficacy: a review. Cancer Res 44:4224–4232 [PubMed] [Google Scholar]
  2. Babson JR, Reed DJ (1978) Inactivation of glutathione reductase by 2-chloroethyl nitrosourea-derived isocyanates. Biochem Biophys Res Commun 83:754–762 [DOI] [PubMed] [Google Scholar]
  3. Bertoncello I, Bradley TR, Webber LM, Hodgson GS, Campbell JJ (1985) Human tumour cell lines established using clonal agar culture. Aust J Exp Biol Med Sci 63:241–248 [DOI] [PubMed] [Google Scholar]
  4. Bonn MH, Parsons PG (1984) Cyclophosphamide resistance developed in a human melanoma cell line. Cancer Treat Rep 68:1239–1246 [PubMed] [Google Scholar]
  5. Constanzi JJ (1983) The chemotherapy of human malignant melanoma. In: Constanzi JJ (ed) Malignant melanoma 1. Nijhoff, The Hague, pp 259–274 [Google Scholar]
  6. Doroshow JH (1983) Effect of anthracycline antibiotics on oxygen radical formation in rat heart. Cancer Res 43:460–472 [PubMed] [Google Scholar]
  7. Dorr RT (1987) Reduced thiol content in L1210 cells treated with BSO increases DNA crosslinking by melphalan. Biochem Biophys Res Commun 144:47–52 [DOI] [PubMed] [Google Scholar]
  8. Dulik DM, Fenselau C (1987) Conversion of melphalan to 4-(glutathionyl)phenylalanine. A novel mechanism for conjugation by glutathione S-transferases. Drug Metab Dispos 15:195–199 [PubMed] [Google Scholar]
  9. Dulik DM, Fenselau C, Hilton J (1986) Characterization of melphalan-glutathione adducts whose formation is catalyzed by glutathione transferases. Biochem Pharmacol 35:3405–3409 [DOI] [PubMed] [Google Scholar]
  10. Erickson LC, Bradley MO, Ducore JM, Ewig RA, Kohn KW (1980) DNA cross-linking and cytotoxicity in normal and transformed human cells treated with antitumor nitrosoureas. Proc Natl Acad Sci USA 77:467–471 [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Feucht KA, Walker MJ, DasGupta TK, Beattie CW (1988) Effect of 17β-estradiol on the growth of estrogen receptor-positive human melanoma in vitro and in athymic mice. Cancer Res 48:7093–7101 [PubMed] [Google Scholar]
  12. Geerarts L, Nathanson L (1986) Non-investigational cytotoxic agents. In: Nathanson L (ed) Contemporary issues in clinical oncology, vol 6. Management of advanced melanoma. Churchill Livingstone, New York, pp 1–32 [Google Scholar]
  13. Green JA, Vistica DT, Young RC, Hamilton TC, Rogan AM, Ozols RF (1984) Potentiation of melphalan cytotoxicity in human ovarian cancer cell lines by glutathione depletion. Cancer Res 44:5427–5431 [PubMed] [Google Scholar]
  14. Griffith OW, Meister A (1979a) Potent and specific inhibition of glutathione synthesis of buthionine sulfoximine (S-n-butyl homocysteine sulfoximine). J Biol Chem 254:7558–7560 [PubMed] [Google Scholar]
  15. Griffith OW, Meister A (1979b) Glutathione: interorgan translocation, turnover, and metabolism. Proc Natl Acad Sci USA 76:5606–5610 [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hamilton TC, Winker MA, Louie KG, Batist G, Behrens BC, Tsuruo T, Grotzinger KP, McKoy WM, Young RC, Ozols RF (1985) 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 34:2583–2586 [DOI] [PubMed] [Google Scholar]
  17. Hansson J, Edgren M, Ehrsson H, Ringborg U, Nilsson B (1988) Effect of D,L-buthionine-5′,R-sulfoximine on cytotoxicity and DNA cross-linking induced by bifunctional DNA-reactive cytostatic drugs in human melanoma cells. Cancer Res 48:19–26 [PubMed] [Google Scholar]
  18. Jordan J, Doherty MD, Cohen GM (1987) Effects of glutathione depletion on the cytotoxicity of agents toward a human colonic tumour cell line. Br J Cancer 55:627–631 [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kappus H, Sies H (1981) Toxic drug effects associated with oxygen metabolism: redox cycling and lipid peroxidation. Experientia 37:1233–1241 [DOI] [PubMed] [Google Scholar]
  20. Kramer RA, Greene K, Ahmed S, Vistica DT (1987) Chemosensitization of L-phenylalanine mustard by the thiol-modulating agent buthionine sulfoximine. Cancer Res 47:1593–1597 [PubMed] [Google Scholar]
  21. Krementz E (1986) Lucy Wortham James lecture. Regional perfusion. Current sophistication, what next? Cancer 57:416–432 [DOI] [PubMed] [Google Scholar]
  22. Levi JA, Dalley DN, Aroney RS (1979) Improved combination chemotherapy in advanced gastric cancer. Br Med J 2:1471–1473 [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Meister A, Anderson ME (1983) Glutathione. Ann Rev Biochem 52:711–760 [DOI] [PubMed] [Google Scholar]
  24. Meredith MJ, Reed DJ (1983) Depletion in vitro of mitochondrial glutathione in rat hepatocytes and enhancement of lipid peroxidation by adriamycin and 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU). Biochem Pharmacol 32:1383–1388 [DOI] [PubMed] [Google Scholar]
  25. Nicotera P, Orrenius S (1986) Role of thiols in protection against biological reactive intermediates. Adv Exp Med Biol 197:41–51 [DOI] [PubMed] [Google Scholar]
  26. Parsons PG, Carter FB, Morrison L, St. Regins M (1981) Mechanism of melphalan resistance developed in vitro in human melanoma cells. Cancer Res 41:1525–1534 [PubMed] [Google Scholar]
  27. Presant CA, Klahr C (1978) Adriamycin, 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU, NSC no. 409962), cyclophosphamide plus prednisone (ABC-P) in melphalan-resistant multiple myeloma. Cancer 42:1222–1227 [DOI] [PubMed] [Google Scholar]
  28. Reed DJ, Babson JR, Beatty PW, Brodie AE, Ellis WW, Potter DW (1980) High-performance liquid chromatography analysis of nanomole levels of glutathione, glutathione disulfide, and related thiols and disulfides. Anal Biochem 106:55–62 [DOI] [PubMed] [Google Scholar]
  29. Richon VM, Schulte N, Eastman A (1987) Multiple mechanisms of resistance to cis-diamminedichloroplatinum(II) in murine leukemia L1210 cells. Cancer Res 47:2056–2061 [PubMed] [Google Scholar]
  30. Russo A, Tochner Z, Philips T, Carmichael J, DeGraff W, Friedman N, Fisher J, Mitchell JB (1986) In vivo modulation of glutathione by buthionine sulfoximine: effect on marrow response to melphalan. Int J Rad Biol Phys 12:1187–1189 [DOI] [PubMed] [Google Scholar]
  31. Skapek SX, Colvin OM, Griffith OW, Elion GB, Bigner DD, Friedman HS (1988) Enhanced melphalan cytotoxicity following buthionine sulfoximine-mediated glutathione depletion in a human medulloblastoma xenograft in athymic mice. Cancer Res 48:2764–2767 [PubMed] [Google Scholar]
  32. Somfai-Relle S, Suzukake K, Vistica BP, Vistica DT (1984) Reduction in cellular glutathione by buthionine sulfoximine and sensitization of murine tumor cells resistant to L-phenylalanine mustard. Biochem Pharmacol 33:485–490 [DOI] [PubMed] [Google Scholar]
  33. Sutow WW, Gehan EA, Dyment PG, Vietti T, Miale T (1978) Multidrug adjuvant chemotherapy for osteosarcoma: interim report of the Southwest Oncology Group Studies. Cancer Treat Rep 62:265–269 [PubMed] [Google Scholar]
  34. Suzukake K, Petro BJ, Vistica DT (1982) Reduction in glutathione content of L-PAM resistant L1210 cells confers drug sensitivity. Biochem Pharmacol 31:121–124 [DOI] [PubMed] [Google Scholar]
  35. Suzukake K, Vistica BP, Vistica DT (1983) Dechlorination of L-phenylalanine mustard by sensitive and resistant tumor cells and its relationship to intracellular glutathione content. Biochem Pharmacol 32:165–167 [DOI] [PubMed] [Google Scholar]
  36. Torti FM, Carter SK (1980) The chemotherapy of prostatic adenocarcinoma. Ann Int Med 92:681–689 [DOI] [PubMed] [Google Scholar]

Articles from Journal of Cancer Research and Clinical Oncology are provided here courtesy of Springer

RESOURCES