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. 1987 Nov;84(21):7668–7671. doi: 10.1073/pnas.84.21.7668

Elevated DNA topoisomerase II activity in nitrogen mustard-resistant human cells.

K B Tan 1, M R Mattern 1, R A Boyce 1, P S Schein 1
PMCID: PMC299361  PMID: 2823270

Abstract

A human Burkitt lymphoma cell line, Raji-HN2, made 10-fold more resistant to nitrogen mustard (HN2) than the parental Raji cell line, exhibited the following characteristics when compared to the parental Raji cells: (i) decreased HN2-induced DNA interstrand crosslinking; (ii) increased (3-fold) DNA topoisomerase II [DNA topoisomerase (ATP-hydrolyzing), EC 5.99.1.3] activity; (iii) increased (4- to 11-fold) sensitivity to topoisomerase II inhibitors; (iv) increased (2-fold) glutathione content; and (v) increased (2-fold) cell doubling time. The resistant phenotype was unstable and was maintained by weekly treatment of the cells with HN2. Growing the resistant cells in the absence of HN2 resulted in a time-dependent decrease in both resistance to HN2 and topoisomerase II activity and an increase in DNA interstrand crosslinking induced by HN2. We hypothesize that HN2 resistance is due to enhanced monoadduct repair with resultant decreased DNA crosslinking and that this process is mediated by topoisomerase II.

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Selected References

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  1. 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]
  2. Berrios M., Osheroff N., Fisher P. A. In situ localization of DNA topoisomerase II, a major polypeptide component of the Drosophila nuclear matrix fraction. Proc Natl Acad Sci U S A. 1985 Jun;82(12):4142–4146. doi: 10.1073/pnas.82.12.4142. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. Day R. S., 3rd, Ziolkowski C. H., Scudiero D. A., Meyer S. A., Lubiniecki A. S., Girardi A. J., Galloway S. M., Bynum G. D. Defective repair of alkylated DNA by human tumour and SV40-transformed human cell strains. Nature. 1980 Dec 25;288(5792):724–727. doi: 10.1038/288724a0. [DOI] [PubMed] [Google Scholar]
  5. DiNardo S., Voelkel K., Sternglanz R. DNA topoisomerase II mutant of Saccharomyces cerevisiae: topoisomerase II is required for segregation of daughter molecules at the termination of DNA replication. Proc Natl Acad Sci U S A. 1984 May;81(9):2616–2620. doi: 10.1073/pnas.81.9.2616. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. ELLMAN G. L. Tissue sulfhydryl groups. Arch Biochem Biophys. 1959 May;82(1):70–77. doi: 10.1016/0003-9861(59)90090-6. [DOI] [PubMed] [Google Scholar]
  7. Earnshaw W. C., Halligan B., Cooke C. A., Heck M. M., Liu L. F. Topoisomerase II is a structural component of mitotic chromosome scaffolds. J Cell Biol. 1985 May;100(5):1706–1715. doi: 10.1083/jcb.100.5.1706. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Erickson L. C., Laurent G., Sharkey N. A., Kohn K. W. DNA cross-linking and monoadduct repair in nitrosourea-treated human tumour cells. Nature. 1980 Dec 25;288(5792):727–729. doi: 10.1038/288727a0. [DOI] [PubMed] [Google Scholar]
  9. Frei E., 3rd, Cucchi C. A., Rosowsky A., Tantravahi R., Bernal S., Ervin T. J., Ruprecht R. M., Haseltine W. A. Alkylating agent resistance: in vitro studies with human cell lines. Proc Natl Acad Sci U S A. 1985 Apr;82(7):2158–2162. doi: 10.1073/pnas.82.7.2158. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. GILMAN A. The initial clinical trial of nitrogen mustard. Am J Surg. 1963 May;105:574–578. doi: 10.1016/0002-9610(63)90232-0. [DOI] [PubMed] [Google Scholar]
  11. Gasser S. M., Laroche T., Falquet J., Boy de la Tour E., Laemmli U. K. Metaphase chromosome structure. Involvement of topoisomerase II. J Mol Biol. 1986 Apr 20;188(4):613–629. doi: 10.1016/s0022-2836(86)80010-9. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Heartlein M. W., Tsuji H., Latt S. A. 5-Bromodeoxyuridine-dependent increase in sister chromatid exchange formation in Bloom's syndrome is associated with reduction in topoisomerase II activity. Exp Cell Res. 1987 Mar;169(1):245–254. doi: 10.1016/0014-4827(87)90242-4. [DOI] [PubMed] [Google Scholar]
  14. Hemminki K., Ludlum D. B. Covalent modification of DNA by antineoplastic agents. J Natl Cancer Inst. 1984 Nov;73(5):1021–1028. [PubMed] [Google Scholar]
  15. Holm C., Goto T., Wang J. C., Botstein D. DNA topoisomerase II is required at the time of mitosis in yeast. Cell. 1985 Jun;41(2):553–563. doi: 10.1016/s0092-8674(85)80028-3. [DOI] [PubMed] [Google Scholar]
  16. Liu L. F., Davis J. L., Calendar R. Novel topologically knotted DNA from bacteriophage P4 capsids: studies with DNA topoisomerases. Nucleic Acids Res. 1981 Aug 25;9(16):3979–3989. doi: 10.1093/nar/9.16.3979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Marx J. L. Drug resistance of cancer cells probed. Science. 1986 Nov 14;234(4778):818–820. doi: 10.1126/science.2877493. [DOI] [PubMed] [Google Scholar]
  18. Miller K. G., Liu L. F., Englund P. T. A homogeneous type II DNA topoisomerase from HeLa cell nuclei. J Biol Chem. 1981 Sep 10;256(17):9334–9339. [PubMed] [Google Scholar]
  19. Ross W. E., Ewig R. A., Kohn K. W. Differences between melphalan and nitrogen mustard in the formation and removal of DNA cross-links. Cancer Res. 1978 Jun;38(6):1502–1506. [PubMed] [Google Scholar]
  20. Russo A., DeGraff W., Friedman N., Mitchell J. B. Selective modulation of glutathione levels in human normal versus tumor cells and subsequent differential response to chemotherapy drugs. Cancer Res. 1986 Jun;46(6):2845–2848. [PubMed] [Google Scholar]
  21. Schaeffer W. I., Friend K. Efficient detection of soft agar grown colonies using a tetrazolium salt. Cancer Lett. 1976 May;1(5):259–262. doi: 10.1016/s0304-3835(75)97506-0. [DOI] [PubMed] [Google Scholar]
  22. Singer B., Brent T. P. Human lymphoblasts contain DNA glycosylase activity excising N-3 and N-7 methyl and ethyl purines but not O6-alkylguanines or 1-alkyladenines. Proc Natl Acad Sci U S A. 1981 Feb;78(2):856–860. doi: 10.1073/pnas.78.2.856. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sullivan D. M., Glisson B. S., Hodges P. K., Smallwood-Kentro S., Ross W. E. Proliferation dependence of topoisomerase II mediated drug action. Biochemistry. 1986 Apr 22;25(8):2248–2256. doi: 10.1021/bi00356a060. [DOI] [PubMed] [Google Scholar]
  24. Tan K. B., Mattern M. R., Boyce R. A., Hertzberg R. P., Schein P. S. Elevated topoisomerase II activity and altered chromatin in nitrogen mustard-resistant human cells. NCI Monogr. 1987;(4):95–98. [PubMed] [Google Scholar]
  25. Taudou G., Mirambeau G., Lavenot C., der Garabedian A., Vermeersch J., Duguet M. DNA topoisomerase activities in concanavalin A-stimulated lymphocytes. FEBS Lett. 1984 Oct 29;176(2):431–435. doi: 10.1016/0014-5793(84)81212-0. [DOI] [PubMed] [Google Scholar]
  26. Teicher B. A., Cucchi C. A., Lee J. B., Flatow J. L., Rosowsky A., Frei E., 3rd Alkylating agents: in vitro studies of cross-resistance patterns in human cell lines. Cancer Res. 1986 Sep;46(9):4379–4383. [PubMed] [Google Scholar]
  27. Vosberg H. P. DNA topoisomerases: enzymes that control DNA conformation. Curr Top Microbiol Immunol. 1985;114:19–102. doi: 10.1007/978-3-642-70227-3_2. [DOI] [PubMed] [Google Scholar]
  28. Wang J. C. DNA topoisomerases. Annu Rev Biochem. 1985;54:665–697. doi: 10.1146/annurev.bi.54.070185.003313. [DOI] [PubMed] [Google Scholar]

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