Skip to main content
NCBI home page
British Journal of Cancer logoLink to British Journal of Cancer
. 1992 Aug;66(2):331–336. doi: 10.1038/bjc.1992.265

Cyclophosphamide decreases O6-alkylguanine-DNA alkyltransferase activity in peripheral lymphocytes of patients undergoing bone marrow transplantation.

S M Lee 1, D Crowther 1, J H Scarffe 1, M Dougal 1, R H Elder 1, J A Rafferty 1, G P Margison 1
PMCID: PMC1977821  PMID: 1387001

Abstract

O6-alkylguanine-DNA-alkyltransferase (ATase) levels were measured in extracts of peripheral blood lymphocytes taken at various times during chemotherapy from 19 patients with various haematological malignancies. Seven patients with advanced Hodgkin's disease received preparative treatment consisting of cyclophosphamide (1.5 g m-2, daily) administered on days 1 to 4 and BCNU (600 mg m-2) on day 5 prior to autologous bone marrow rescue (ABMR) delivered on day 7. Treatment in the remaining 12 patients consisted of cyclophosphamide (1.8 g m-2, daily) given on days 1 and 2 followed at day 4 with total body irradiation (TBI) administered in six fractions over the subsequent 3 days to a total dose of 1200 cGy prior to bone marrow transplantation. In the Hodgkin's group, significant decreases in ATase activity were seen during the cyclophosphamide treatment, and the median ATase nadir was 32% (range 0% to 57%) of pretreatment levels following 4 days of cyclophosphamide. In one patient, no ATase activity was detectable following the 4th cyclophosphamide treatment. ATase activities decreased further after BCNU administration to a median of 19% (range 0% to 32%) of pretreatment levels. Extensive cyclophosphamide-induced reduction of lymphocyte ATase levels was also seen in the other group of 12 patients treated with cyclophosphamide/TBI: postcyclophosphamide median ATase nadir was 35% (range 12% to 78%) of the pretreatment levels. No ATase depletion was seen when cyclophosphamide (up to 10 mM) was incubated for 2 h with pure recombinant human ATase in vitro whereas ATase activity was reduced by 90% on preincubation with 100 microns acrolein or with greater than 1 mM phosphoramide mustard. This suggests that a cyclophosphamide-induced decrease in ATase levels in human peripheral lymphocytes in vivo may be due to depletion mediated by the production of intracellular acrolein. Since ATase appears to be a principal mechanism in cellular resistance to the cytotoxic effects of BCNU and related alkylating agents, these observations suggest that a cyclophosphamide-induced reduction in ATase activity may be an additional factor in the effectiveness of the combined sequential therapy.

Full text

PDF
331

Selected References

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

  1. Ahmed T., Ciavarella D., Feldman E., Ascensao J., Hussain F., Engelking C., Gingrich S., Mittelman A., Coleman M., Arlin Z. A. High-dose, potentially myeloablative chemotherapy and autologous bone marrow transplantation for patients with advanced Hodgkin's disease. Leukemia. 1989 Jan;3(1):19–22. [PubMed] [Google Scholar]
  2. Ali-Osman F., Caughlan J., Gray G. S. Decreased DNA interstrand cross-linking and cytotoxicity induced in human brain tumor cells by 1,3-bis(2-chloroethyl)-1-nitrosourea after in vitro reaction with glutathione. Cancer Res. 1989 Nov 1;49(21):5954–5958. [PubMed] [Google Scholar]
  3. Armitage J. O., Barnett M. J., Carella A. M., Dicke K. A., Diehl V., Gribben J. G., Pfreundschuh M. Bone marrow transplantation in the treatment of Hodgkin's lymphoma: problems, remaining challenges, and future prospects. Recent Results Cancer Res. 1989;117:246–253. doi: 10.1007/978-3-642-83781-4_28. [DOI] [PubMed] [Google Scholar]
  4. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  5. Brennand J., Margison G. P. Reduction of the toxicity and mutagenicity of alkylating agents in mammalian cells harboring the Escherichia coli alkyltransferase gene. Proc Natl Acad Sci U S A. 1986 Sep;83(17):6292–6296. doi: 10.1073/pnas.83.17.6292. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brent T. P. Isolation and purification of O6-alkylguanine-DNA alkyltransferase from human leukemic cells. Prevention of chloroethylnitrosourea-induced cross-links by purified enzyme. Pharmacol Ther. 1985;31(1-2):121–140. doi: 10.1016/0163-7258(85)90040-3. [DOI] [PubMed] [Google Scholar]
  7. Brock N. Oxazaphosphorine cytostatics: past-present-future. Seventh Cain Memorial Award lecture. Cancer Res. 1989 Jan 1;49(1):1–7. [PubMed] [Google Scholar]
  8. Carella A. M., Congiu A. M., Gaozza E., Mazza P., Ricci P., Visani G., Meloni G., Cimino G., Mangoni L., Coser P. High-dose chemotherapy with autologous bone marrow transplantation in 50 advanced resistant Hodgkin's disease patients: an Italian study group report. J Clin Oncol. 1988 Sep;6(9):1411–1416. doi: 10.1200/JCO.1988.6.9.1411. [DOI] [PubMed] [Google Scholar]
  9. Carmichael J., Adams D. J., Ansell J., Wolf C. R. Glutathione and glutathione transferase levels in mouse granulocytes following cyclophosphamide administration. Cancer Res. 1986 Feb;46(2):735–739. [PubMed] [Google Scholar]
  10. Chresta C. M., Crook T. R., Souhami R. L. Depletion of cellular glutathione by N,N'-bis(trans-4-hydroxycyclohexyl)-N'-nitrosourea as a determinant of sensitivity of K562 human leukemia cells to 4-hydroperoxycyclophosphamide. Cancer Res. 1990 Jul 1;50(13):4067–4071. [PubMed] [Google Scholar]
  11. D'Incalci M., Citti L., Taverna P., Catapano C. V. Importance of the DNA repair enzyme O6-alkyl guanine alkyltransferase (AT) in cancer chemotherapy. Cancer Treat Rev. 1988 Dec;15(4):279–292. doi: 10.1016/0305-7372(88)90026-6. [DOI] [PubMed] [Google Scholar]
  12. Dolan M. E., Corsico C. D., Pegg A. E. Exposure of HeLa cells to 0(6)-alkylguanines increases sensitivity to the cytotoxic effects of alkylating agents. Biochem Biophys Res Commun. 1985 Oct 15;132(1):178–185. doi: 10.1016/0006-291x(85)91004-6. [DOI] [PubMed] [Google Scholar]
  13. Dolan M. E., Mitchell R. B., Mummert C., Moschel R. C., Pegg A. E. Effect of O6-benzylguanine analogues on sensitivity of human tumor cells to the cytotoxic effects of alkylating agents. Cancer Res. 1991 Jul 1;51(13):3367–3372. [PubMed] [Google Scholar]
  14. Dolan M. E., Moschel R. C., Pegg A. E. Depletion of mammalian O6-alkylguanine-DNA alkyltransferase activity by O6-benzylguanine provides a means to evaluate the role of this protein in protection against carcinogenic and therapeutic alkylating agents. Proc Natl Acad Sci U S A. 1990 Jul;87(14):5368–5372. doi: 10.1073/pnas.87.14.5368. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Draeger J., Peter G., Hohorst H. J. Deactivation of cyclophosphamide (NSC-26271) metabolites by sulfhydryl compounds. Cancer Treat Rep. 1976 Apr;60(4):355–359. [PubMed] [Google Scholar]
  16. Futscher B. W., Micetich K. C., Barnes D. M., Fisher R. I., Erickson L. C. Inhibition of a specific DNA repair system and nitrosourea cytotoxicity in resistant human cancer cells. Cancer Commun. 1989;1(1):65–73. doi: 10.3727/095535489820875444. [DOI] [PubMed] [Google Scholar]
  17. Gerson S. L., Miller K., Berger N. A. O6 alkylguanine-DNA alkyltransferase activity in human myeloid cells. J Clin Invest. 1985 Dec;76(6):2106–2114. doi: 10.1172/JCI112215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Gerson S. L. Modulation of human lymphocyte O6-alkylguanine-DNA alkyltransferase by streptozotocin in vivo. Cancer Res. 1989 Jun 1;49(11):3134–3138. [PubMed] [Google Scholar]
  19. Gerson S. L., Trey J. E., Miller K. Potentiation of nitrosourea cytotoxicity in human leukemic cells by inactivation of O6-alkylguanine-DNA alkyltransferase. Cancer Res. 1988 Mar 15;48(6):1521–1527. [PubMed] [Google Scholar]
  20. Gingrich R. D., Ginder G. D., Burns L. J., Wen B. C., Fyfe M. A. BVAC ablative chemotherapy followed by autologous bone marrow transplantation for patients with advanced lymphoma. Blood. 1990 Jun 15;75(12):2276–2281. [PubMed] [Google Scholar]
  21. Gonzaga P. E., Harris L., Margison G. P., Brent T. P. Evidence that covalent complex formation between BCNU-treated oligonucleotides and E. coli alkyltransferases requires the O6-alkylguanine function. Nucleic Acids Res. 1990 Jul 11;18(13):3961–3966. doi: 10.1093/nar/18.13.3961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Gribben J. G., Linch D. C., Singer C. R., McMillan A. K., Jarrett M., Goldstone A. H. Successful treatment of refractory Hodgkin's disease by high-dose combination chemotherapy and autologous bone marrow transplantation. Blood. 1989 Jan;73(1):340–344. [PubMed] [Google Scholar]
  23. Gurtoo H. L., Hipkens J. H., Sharma S. D. Role of glutathione in the metabolism-dependent toxicity and chemotherapy of cyclophosphamide. Cancer Res. 1981 Sep;41(9 Pt 1):3584–3591. [PubMed] [Google Scholar]
  24. Herrmann F., Sieber G., Jauer B., Lochner A., Komischke B., Rühl H. Evaluation of the circulating and splenic lymphocyte subpopulations in patients with non-Hodgkin lymphomas and Hodgkin's disease using monoclonal antibodies. Blut. 1983 Jul;47(1):41–51. doi: 10.1007/BF00321049. [DOI] [PubMed] [Google Scholar]
  25. Jardine I., Fenselau C., Appler M., Kan M. N., Brundrett R. B., Colvin M. Quantitation by gas chromatography-chemical ionization mass spectrometry of cyclophosphamide, phosphoramide mustard, and nornitrogen mustard in the plasma and urine of patients receiving cyclophosphamide therapy. Cancer Res. 1978 Feb;38(2):408–415. [PubMed] [Google Scholar]
  26. Juma F. D., Rogers H. J., Trounce J. R. Pharmacokinetics of cyclophosphamide and alkylating activity in man after intravenous and oral administration. Br J Clin Pharmacol. 1979 Sep;8(3):209–217. doi: 10.1111/j.1365-2125.1979.tb01004.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kleihues P., Margison G. P. Exhaustion and recovery of repair excision of O6-methylguanine from rat liver DNA. Nature. 1976 Jan 15;259(5539):153–155. doi: 10.1038/259153a0. [DOI] [PubMed] [Google Scholar]
  28. Lee F. Y., Flannery D. J., Siemann D. W. Prediction of tumour sensitivity to 4-hydroperoxycyclophosphamide by a glutathione-targeted assay. Br J Cancer. 1991 Feb;63(2):217–222. doi: 10.1038/bjc.1991.52. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Lee F. Y. Glutathione diminishes the anti-tumour activity of 4-hydroperoxycyclophosphamide by stabilising its spontaneous breakdown to alkylating metabolites. Br J Cancer. 1991 Jan;63(1):45–50. doi: 10.1038/bjc.1991.10. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Lee S. M., Thatcher N., Margison G. P. O6-alkylguanine-DNA alkyltransferase depletion and regeneration in human peripheral lymphocytes following dacarbazine and fotemustine. Cancer Res. 1991 Jan 15;51(2):619–623. [PubMed] [Google Scholar]
  31. Margison G. P., Butler J., Hoey B. O6-Methylguanine methyltransferase activity is increased in rat tissues by ionising radiation. Carcinogenesis. 1985 Dec;6(12):1699–1702. doi: 10.1093/carcin/6.12.1699. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. Meer L., Schold S. C., Kleihues P. Inhibition of the hepatic O6-alkylguanine-DNA alkyltransferase in vivo by pretreatment with antineoplastic agents. Biochem Pharmacol. 1989 Mar 15;38(6):929–934. doi: 10.1016/0006-2952(89)90282-7. [DOI] [PubMed] [Google Scholar]
  34. Pegg A. E. Mammalian O6-alkylguanine-DNA alkyltransferase: regulation and importance in response to alkylating carcinogenic and therapeutic agents. Cancer Res. 1990 Oct 1;50(19):6119–6129. [PubMed] [Google Scholar]
  35. Reece D. E., Barnett M. J., Connors J. M., Fairey R. N., Fay J. W., Greer J. P., Herzig G. P., Herzig R. H., Klingemann H. G., LeMaistre C. F. Intensive chemotherapy with cyclophosphamide, carmustine, and etoposide followed by autologous bone marrow transplantation for relapsed Hodgkin's disease. J Clin Oncol. 1991 Oct;9(10):1871–1879. doi: 10.1200/JCO.1991.9.10.1871. [DOI] [PubMed] [Google Scholar]
  36. Sagher D., Karrison T., Schwartz J. L., Larson R., Meier P., Strauss B. Low O6-alkylguanine DNA alkyltransferase activity in the peripheral blood lymphocytes of patients with therapy-related acute nonlymphocytic leukemia. Cancer Res. 1988 Jun 1;48(11):3084–3089. [PubMed] [Google Scholar]
  37. Santibanez-Koref M., Elder R. H., Fan C. Y., Cawkwell L., McKie J. H., Douglas K. T., Margison G. P., Rafferty J. A. Isolation and partial characterization of murine O6-alkylguanine-DNA-alkyltransferase: comparative sequence and structural properties. Mol Carcinog. 1992;5(2):161–169. doi: 10.1002/mc.2940050212. [DOI] [PubMed] [Google Scholar]
  38. Sladek N. E., Doeden D., Powers J. F., Krivit W. Plasma concentrations of 4-hydroxycyclophosphamide and phosphoramide mustard in patients repeatedly given high doses of cyclophosphamide in preparation for bone marrow transplantation. Cancer Treat Rep. 1984 Oct;68(10):1247–1254. [PubMed] [Google Scholar]
  39. Spitzer G., Dicke K. A., Litam J., Verma D. S., Zander A., Lanzotti V., Valdivieso M., McCredie K. B., Samuels M. L. High-dose combination chemotherapy with autologous bone marrow transplantation in adult solid tumors. Cancer. 1980 Jun 15;45(12):3075–3085. doi: 10.1002/1097-0142(19800615)45:12<3075::aid-cncr2820451233>3.0.co;2-8. [DOI] [PubMed] [Google Scholar]
  40. Stammberger I., Schmahl W., Nice L. The effects of X-irradiation, N-ethyl-N-nitrosourea or combined treatment on O6-alkylguanine-DNA alkyltransferase activity in fetal rat brain and liver and the induction of CNS tumours. Carcinogenesis. 1990 Feb;11(2):219–222. doi: 10.1093/carcin/11.2.219. [DOI] [PubMed] [Google Scholar]
  41. Tong W. P., Kirk M. C., Ludlum D. B. Formation of the cross-link 1-[N3-deoxycytidyl),2-[N1-deoxyguanosinyl]ethane in DNA treated with N,N'-bis(2-chloroethyl)-N-nitrosourea. Cancer Res. 1982 Aug;42(8):3102–3105. [PubMed] [Google Scholar]
  42. Yarosh D. B., Hurst-Calderone S., Babich M. A., Day R. S., 3rd Inactivation of O6-methylguanine-DNA methyltransferase and sensitization of human tumor cells to killing by chloroethylnitrosourea by O6-methylguanine as a free base. Cancer Res. 1986 Apr;46(4 Pt 1):1663–1668. [PubMed] [Google Scholar]
  43. Zlotogorski C., Erickson L. C. Pretreatment of human colon tumor cells with DNA methylating agents inhibits their ability to repair chloroethyl monoadducts. Carcinogenesis. 1984 Jan;5(1):83–87. doi: 10.1093/carcin/5.1.83. [DOI] [PubMed] [Google Scholar]

Articles from British Journal of Cancer are provided here courtesy of Cancer Research UK

RESOURCES