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Role of mismatch repair in the induction of chromosomal aberrations and sister chromatid exchanges in cells treated with different chemotherapeutic agents

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Abstract

Purpose

The mismatch repair (MMR) system plays a major role in mediating the cytotoxicity and clastogenicity of agents generating O6-methylguanine in DNA. Loss of MMR has also been associated with tumor cell resistance to the cytotoxic effects of 6-thioguanine and cisplatin and with hypersensitivity to N-(2-chloroethyl)-N′-cyclohexyl-N-nitrosourea (CCNU). The aim of the present investigation was to elucidate the role played by the MMR system in the generation of chromosomal damage in cells exposed to 6-thioguanine, cisplatin or CCNU.

Methods

The MMR-proficient cell lines TK6 and HCT116/3-6, and their MMR-deficient counterparts MT1 and HCT116, were treated with 6-thioguanine, cisplatin or CCNU, and analyzed for cell growth inhibition and chromosomal damage. As a control, similar experiments were performed with the methylating agent temozolomide.

Results

Cytotoxicity, chromosomal aberrations and sister chromatid exchanges induced by 6-thioguanine and temozolomide were significantly reduced in the MMR-deficient cell lines with respect to their MMR-proficient counterparts. In contrast, although conferring some protection against cytotoxicity, the loss of MMR did not affect cytogenetic damage induced by cisplatin. CCNU produced comparable levels of cytotoxicity, chromosomal aberrations and sister chromatid exchanges in both MMR-proficient and MMR-deficient cell lines.

Conclusions

The MMR system is involved in the generation of chromosomal damage in cells exposed to 6-thioguanine. The system does not play a relevant role in the generation of chromosomal damage in cells treated with CDDP and does not confer protection against the clastogenic effects of CCNU, at least in the cell lines investigated.

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Fig. 1a–d.
Fig. 2a–d.
Fig. 3a, b.

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Abbreviations

BG:

O6-Benzylguanine

CCNU:

N-(2-Chloroethyl)-N′-cyclohexyl-N-nitrosourea

CDDP:

cis-Diamino-di-chloro-platinum (II)

MGMT:

O6-Methylguanine-DNA methyltransferase

MMR:

Mismatch repair

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

sce:

Sister chromatid exchanges

6-TG:

6-Thioguanine

TMZ (temozolomide):

8-Carbamoyl-3-methyl-imidazo[5,1-d]-1,2,3,5-tetrazin-4(3H)-one

References

  1. Aebi S, Fink D, Gordon R, Kim HK, Zheng H, Fink JL, Howell SB (1997) Resistance to cytotoxic drugs in DNA mismatch repair-deficient cells. Clin Cancer Res 3:1763

    PubMed  Google Scholar 

  2. Allan MJ, Engelward BP, Dreslin AJ, Wyatt MD, Tomasz M, Samson LD (1998) Mammalian 3-methyladenine DNA glycosylase protects against the toxicity and clastogenicity of certain chemotherapeutic DNA cross-linking agents. Cancer Res 58:3965

    CAS  PubMed  Google Scholar 

  3. Aquilina G, Ceccotti S, Martinelli S, Hampson R, Bignami M (1998) N-(2-chloroethyl)-N′-cyclohexyl-N-nitrosurea sensitivity in mismatch repair-defective human cells. Cancer Res 58:135

    CAS  PubMed  Google Scholar 

  4. Armstrong MJ, Galloway SM (1997) Mismatch repair provokes chromosome aberrations in hamster cells treated with methylating agents or 6-thioguanine, but not with ethylating agents. Mutat Res 373:167

    Article  CAS  PubMed  Google Scholar 

  5. Bean CL, Bradt CI, Hill R, Johnson TE, Stallworth MV, Galloway SM (1994) Chromosome abs: persistence of alkylation damage and modulation of O6-alkylguanine-DNA alkyltransferase. Mutat Res 307:67

    Article  CAS  PubMed  Google Scholar 

  6. Beranek DT (1990) Distribution of methyl and ethyl adducts following alkylation with monofunctional alkylating agents. Mutat Res 231:11

    Article  CAS  PubMed  Google Scholar 

  7. Bodell WJ, Aida T, Berger MS, Rosenblum ML (1986) Increased repair of 1 O6-alkylguanine DNA adducts in glioma-derived human cells resistant to the cytotoxic and cytogenetic effects of 1,3-bis(2-chloroethyl)-1-nitrosurea. Carcinogenesis 7:879

    CAS  PubMed  Google Scholar 

  8. D'Atri S, Tentori L, Lacal PM, Graziani G, Pagani E, Benincasa E, Zambruno G, Bonmassar E, Jiricny J (1998) Involvement of the mismatch repair system in temozolomide-induced apoptosis. Mol Pharmacol 54:334

    CAS  PubMed  Google Scholar 

  9. D'Incalci M, Citti L, Taverna P, Catapano CV (1988) Importance of DNA repair enzyme O6-alkyltransferase (AT) in cancer chemotherapy. Cancer Treat Rev 15:279

    CAS  PubMed  Google Scholar 

  10. Dolan EM, Moshel RC, Pegg AE (1990) Depletion of 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 87:5368

    CAS  PubMed  Google Scholar 

  11. Duckett DR, Drummond JT, Murchie AI, Reardon JT, Sancar A, Lilley DM, Modrich P (1996) Human MutSα recognizes damaged DNA base pairs containing O-6-methylguanine O-4-methylthymine, or the cisplatin-d (GpG) adduct. Proc Natl Acad Sci U S A 93:6443

    CAS  PubMed  Google Scholar 

  12. Durant ST, Morris MM, Illand M, McKay HJ, McKormick C, Hirst GL, Borts RH, Brown R (1999) Dependence on RAD52 and RAD1 for anticancer drug resistance mediated by inactivation of mismatch repair genes. Curr Biol 9:51

    CAS  PubMed  Google Scholar 

  13. Fink D, Nebel S, Aebi S, Zheng H, Cenni B, Nehmè A, Christen RD, Howell SB (1996) The role of DNA mismatch repair in platinum drug resistance. Cancer Res 56:4881

    CAS  PubMed  Google Scholar 

  14. Fink D, Aebi S, Howell SB (1998) The role of DNA mismatch repair in drug resistance. Clin Cancer Res 4:1

    CAS  PubMed  Google Scholar 

  15. Fiumicino S, Martinelli S, Colussi C, Aquilina G, Leonetti C, Crescenzi M, Bignami M (2000) Sensitivity to DNA cross-linking chemotherapeutic agents in mismatch repair-defective cells in vitro and in xenografts. Int J Cancer 85:590

    Article  CAS  PubMed  Google Scholar 

  16. Galloway SM, Greenwood SK, Hill R, Bradt CI, Bean CL (1995) A role for mismatch repair in production of chromosome aberrations by methylating agents in human cells. Mutat Res 346:231

    Article  CAS  PubMed  Google Scholar 

  17. Goldmacher VS, Cuzick RA, Thilly WG (1986) Isolation and partial characterization of human cell mutants differing in sensitivity to killing and mutation by methylnitrosurea and N-methyl-N′-nitro-N-nitrosoguanidine. J Biol Chem 261:12462

    CAS  PubMed  Google Scholar 

  18. Hansen MB, Nielsen SE, Berg K (1989) Reexamination and further development of a precise and rapid dye method for measuring cell growth/cell kill. J Immunol Methods 119:203

    CAS  PubMed  Google Scholar 

  19. Hawn MT, Umar A, Carethers JM, Marra G, Kunkel TA, Boland CR, Koi M (1995) Evidence for a connection between the mismatch repair system and the G2 cell cycle checkpoint. Cancer Res 55:3721

    CAS  PubMed  Google Scholar 

  20. Jiricny J (1998) Eukaryotic mismatch repair: an update. Mutat Res 409:107

    CAS  PubMed  Google Scholar 

  21. Jiricny J, Nystrom-Lahti M (2000) Mismatch repair defects in cancer. Curr Opin Genet Dev 10:157

    CAS  PubMed  Google Scholar 

  22. Kaina B, Ziouta A, Ochs K, Coquerelle T (1997) Chromosomal instability, reproductive cell death and apoptosis induced by O6-methyl-guanine in Mex−, Mex+ and methylation-tolerant mismatch repair compromised cells: facts and models. Mutat Res 381:227

    Article  CAS  PubMed  Google Scholar 

  23. Karran P (2001) Mechanisms of tolerance to DNA damaging therapeutic drugs. Carcinogenesis 22:1931

    Article  CAS  PubMed  Google Scholar 

  24. Karran P, Bignami M (1992) Self-destruction and tolerance in resistance of mammalian cells to alkylation damage. Nucleic Acids Res 20:2933

    CAS  PubMed  Google Scholar 

  25. Kat A, Thilly WG, Fang WH, Longley MJ, Li GM, Modrich P (1993) An alkylation-tolerant mutator human cell line is deficient in strand-specific mismatch repair. Proc Natl Acad Sci U S A 90:6424

    CAS  PubMed  Google Scholar 

  26. Koi M, Umar A, Chauhan DP, Cherian SP, Carethers JM, Kunkel TA, Boland CR (1994) Human chromosome 3 corrects mismatch repair deficiency and microsatellite instability and reduces N-methyl-N′-nitrosoguanidine tolerance in colon tumor cells with homozygous hMLH1 mutation. Cancer Res 54:4308

    CAS  PubMed  Google Scholar 

  27. Liu L, Markowitz S, Gerson SL (1996) Mismatch repair mutations override alkyltransferase in conferring resistance to temozolomide but not to 1,3-bis (2-chloroethyl)nitrosourea. Cancer Res 56:5375

    CAS  PubMed  Google Scholar 

  28. Ludlum DB (1990) DNA alkylation by haloethylnitrosureas: nature of modifications produced and their enzymatic repair or removal. Mutat Res 233:117

    Article  CAS  PubMed  Google Scholar 

  29. Mello JA, Acharya S, Fishel R, Essingmann JM (1996) The mismatch-repair protein hMSH2 binds selectively to DNA adducts of the anticancer drug cisplatin. Chem Biol 3:579

    CAS  PubMed  Google Scholar 

  30. Modrich P (1997) Strand-specific mismatch repair in mammalian cells. J Biol Chem 272:24727

    Article  CAS  PubMed  Google Scholar 

  31. Papadopoulos N, Nicoladesi NC, Wei YF, Ruben SM, Carte KC, Rosen CA, Haseltine WA, Fleishmann RD, Fraser CM, Adams MD, Venter JC, Hamilton SR, Petersen GM, Watson P, Lynch HT, Peltomaki P, Mecklin JP, de la Chapelle A, Kinzler KW, Vogelstein B (1994) Mutation of a mutL homolog in hereditary colon cancer. Science 263:1625

    CAS  PubMed  Google Scholar 

  32. Pegg AE, Dolan ME, Moschel RC (1995) Structure, function, and inhibition of O6-alkylguanine-DNA alkyltransferase. Prog Nucleic Acid Res Mol Biol 51:167

    CAS  PubMed  Google Scholar 

  33. Perry P, Wolff S (1974) New Giemsa method for the differential staining of sister chromatids. Nature 251:156

    CAS  PubMed  Google Scholar 

  34. Roberts JJ, Friedlos F (1987) Quantitative estimation of cisplatinum induced DNA interstrand cross-links and their repair in mammalian cells: relationship to toxicity. Pharmacol Ther 34:215

    Article  CAS  PubMed  Google Scholar 

  35. Robins P, Harris AL, Lindahl T (1983) Cross-linking of DNA induced by chloroethylnitrosurea is prevented by O6-methylguanine-DNA methyltransferase. Nucleic Acids Res 22:7743

    Google Scholar 

  36. Skopek TR, Liber HL, Penman BW, Thilly WG (1978) Isolation of a human lymphoblastoid line heterozygous at the thymidine kinase locus: possibility for a rapid human cell mutation assay. Biochem Biophys Res Commun 84:411

    CAS  PubMed  Google Scholar 

  37. Swann PF, Waters TR, Moulton DC, Xu YZ, Zheng Q, Edwards M, Mace R (1996) Role of postreplicative DNA mismatch repair in the cytotoxic action of thioguanine. Science 273:1109

    CAS  PubMed  Google Scholar 

  38. Vaisman A, Varchenko M, Umar A, Kunkel TA, Risinger JI, Barrett JC, Hamilton TC, Chaney SG (1998) The role of hMLH1, hMSH3, and hMSH6 defects in cisplatin and oxaliplatin resistance: correlation with replicative bypass of platinum-DNA adducts. Cancer Res 58:3579

    CAS  PubMed  Google Scholar 

  39. Vernole P, Tedeschi B, Caporossi D, Maccarrone M, Melino G, Annicchiarico-Petruzzelli M (1998) Induction of apoptosis by bleomycin in resting and cycling human lymphocytes. Mutagenesis 13:209

    CAS  PubMed  Google Scholar 

  40. Watson AJ, Margison GP (1999) O6-Alkylguanine-DNA alkyltransferase assays. In: Brown R, Boeger-Brown U (eds) Cytotoxic drug resistance mechanisms. Humana Press, Totowa, p 167

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Acknowledgements

The authors thank Prof. E. Bonmassar for his valuable advice, and Mr. G. Bonelli for his skilful technical assistance. The research was partly supported by grants from M.U.R.S.T to P.V., and partly by the Italian Ministry of Health.

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Authors and Affiliations

  1. Department of Public Health and Cell Biology, University of Rome "Tor Vergata", Via Montpellier 1, 00133, Rome, Italy

    Patrizia Vernole

  2. Laboratory of Pharmacology, Istituto Dermopatico dell'Immacolata (IDI-IRCCS), Via dei Monti di Creta 104, 00167, Rome, Italy

    Rita Pepponi & Stefania D'Atri

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  1. Patrizia Vernole
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  2. Rita Pepponi
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Correspondence to Patrizia Vernole.

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Vernole, P., Pepponi, R. & D'Atri, S. Role of mismatch repair in the induction of chromosomal aberrations and sister chromatid exchanges in cells treated with different chemotherapeutic agents. Cancer Chemother Pharmacol 52, 185–192 (2003). https://doi.org/10.1007/s00280-003-0660-6

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