Mutagen-Induced Chromatid Breakage as a Marker of Cancer Risk

  • Xifeng Wu
  • Yun-Ling Zheng
  • T. C. Hsu
Part of the Methods in Molecular Biology™ book series (MIMB, volume 291)


Risk assessment is now recognized as a multidisciplinary process, extending beyond the scope of traditional epidemiologic methodology to include biological evaluation of interindividual differences in carcinogenic susceptibility. Modulation of environmental exposures by host genetic factors may explain much of the observed interindividual variation in susceptibility to carcinogenesis. These genetic factors include, but are not limited to, carcinogen metabolism and DNA repair capacity. This chapter describes a standardized method for the functional assessment of mutagen sensitivity. This in vitro assay measures the frequency of mutagen-induced breaks in peripheral lymphocytes. Mutagen sensitivity assessed by this method has been shown to be a significant risk factor for tobacco-related malignancies, especially those of the upper aerodigestive tract. Mutagen sensitivity may therefore be a useful member of a panel of susceptibility markers for defining high-risk subgroups for chemoprevention trials. This chapter describes methods for and discusses results from studies of mutagen sensitivity as measured by quantifying chromatid breaks induced by chromosome breaking agents, such as the γ-radiation radiomimetic DNA crosslinking agent bleomycin and chemicals that form so-called bulky DNA adducts, such as 4-NQO and the tobacco smoke constituent, benzo[a]pyrene, in short-term cultured peripheral blood lymphocytes.

Key Words

Mutagen sensitivity chromatid breaks cancer susceptibility bleomycin benzo[a]pyrene nitroquinoline γ-irradiation 


  1. 1.
    Hsu, T. C. (1983) Genetic instability in the human population: a working hypothesis. Hereditas 98, 1–9.PubMedCrossRefGoogle Scholar
  2. 2.
    Hsu, T. C, Johnston, D. A., Cherry, L. M., et al. (1989) Sensitivity to genotoxic effects of bleomycin in humans: possible relationship to environmental carcinogenesis. Int. J. Cancer 43, 403–409.PubMedCrossRefGoogle Scholar
  3. 3.
    Hsu, T. C, Spitz, M. R., and Schantz, S. P. (1991) Mutagen sensitivity: a biologic marker of cancer susceptibility. Cancer Epidemiol. Biomarkers Prev. 1, 83–89.PubMedGoogle Scholar
  4. 4.
    Spitz, M. R., Fueger, J. J., Beddingfield, N. A., et al. (1989) Chromosome sensitivity to bleomycin-induced mutagenesis, an independent risk factor for upper aerodigestive tract cancers. Cancer Res. 49, 4626–4628.PubMedGoogle Scholar
  5. 5.
    Schantz, S. P., Hsu, T. C, Ainslie, N., and Moser, R. P. (1989) Young adults with head and neck cancer express increased susceptibility to mutagen-induced chromosome damage. JAMA 262, 3313–3315.PubMedCrossRefGoogle Scholar
  6. 6.
    Spitz, M. R., Fueger, J. J., Halabi, S., Schantz, S. P., Sample, D., and Hsu, T. C. (1993) Mutagen sensitivity in upper aerodigestive tract cancer: a case-control analysis. Cancer Epidemiol. Biomarkers Prev. 2, 329–333.PubMedGoogle Scholar
  7. 7.
    Cloos, J., Steen, I., Joenje, H., et al. (1993) Association between bleomycin genotoxicity and non-constitutional risk factors for head and neck cancer. Cancer Lett. 74, 161–165.PubMedCrossRefGoogle Scholar
  8. 8.
    Cloos, J., Spitz, M. R., Schantz, S. P., et al. (1996) Genetic susceptibility to head and neck squamous cell carcinoma. J. Natl. Cancer Inst. 88, 530–535.PubMedCrossRefGoogle Scholar
  9. 9.
    Wu, X., Gu, J., Hong, W. K., et al. (1998) Benzo[a]pyrene diol epoxide and bleomycin sensitivity and susceptibility to cancer of upper aerodigestive tract. J. Natl. Cancer Inst. 90, 1393–1399.PubMedCrossRefGoogle Scholar
  10. 10.
    Spitz, M. R., Hsu, T. C, Wu, X. F., Fueger, J. J., Amos, C. I., and Roth, J. A. (1995) Mutagen sensitivity as a biologic marker of lung cancer risk in African Americans. Cancer Epidemiol. Biomarkers Prev. 4, 99–103.PubMedGoogle Scholar
  11. 11.
    Strom, S. S., Wu, X., Sigurdson, A. J., et al. (1995) Lung cancer, smoking patterns, and mutagen sensitivity in Mexican-Americans. J. Natl. Cancer Inst. 18, 29–33.Google Scholar
  12. 12.
    Wu, X., Delclos, G. L., Annegers, F. J., et al. (1995) A case-control study of wood-dust exposure, mutagen sensitivity, and lung-cancer risk. Cancer Epidemiol. Biomarkers Prev. 4, 583–588.PubMedGoogle Scholar
  13. 13.
    Wu, X., Gu, J., Amos, C. I., Jiang, H., Hong, W. K., and Spitz, M. R. (1998) A parallel study of in vitro sensitivity to benzo[a]pyrene diol epoxide and bleomycin in lung cancer cases and controls. Cancer 83, 1118–1127.PubMedCrossRefGoogle Scholar
  14. 14.
    Wu, X., Gu, J., Patt, Y., et al. (1998) Mutagen sensitivity as a susceptibility marker for human hepatocellular carcinoma. Cancer Epidemiol. Biomarkers Prev. 7, 567–570.PubMedGoogle Scholar
  15. 15.
    Spitz, M. R., Lippman, S. M., Jiang, H., et al. (1998) Mutagen sensitivity as a predictor of tumor recurrence in patients with cancer of the upper aerodigestive tract. J. Natl. Cancer Inst. 90, 243–245.PubMedCrossRefGoogle Scholar
  16. 16.
    Hsu, T. C, Feun, L., Trizna, Z., et al. (1993) Differential sensitivity among three human subpopulations in response to 4-nitroquinoline-1-oxide and to bleomycin. Int. J. Oncol. 3, 827–830.Google Scholar
  17. 17.
    Bondy, M. L., Kyritis, A. P., Gu, J., et al. (1996) Mutagen sensitivity and risk of glioma: a case-control analysis. Cancer Res. 56, 1484–1486.PubMedGoogle Scholar
  18. 18.
    Buchholz, T. A. and Wu, X. F. (2001) Radiation-induced chromatid breaks as a predictor of breast cancer risk. Int. J. Radiat. Oncol. Biol. Phys. 49, 533–537.PubMedCrossRefGoogle Scholar
  19. 19.
    Wu, X., Hsu, T. C, and Spitz, M. R. (1996) Mutagen sensitivity exhibits a dose-response relationship in case-control studies. Cancer Epidemiol. Biomarkers Prev. 5, 577–578.PubMedGoogle Scholar
  20. 20.
    Xu, Y. J., Kim, E. Y., and Demple, B. (1998) Excision of C-4′-oxidized deoxyribose lesions from double-stranded DNA by human apurinic/apyrimidinic endonuclease (Ape1 protein) and DNA polymerase beta. J. Biol. Chem. 273, 28837–28844.PubMedCrossRefGoogle Scholar
  21. 21.
    Dar, M. E., Winters, T. A., and Jorgensen, T. J. (1997) Identification of defective illegitimate recombinational repair of oxidatively-induced DNA double-strand breaks in ataxia-telangiectasia cells. Mutat. Res. 384, 169–179.PubMedGoogle Scholar
  22. 22.
    Arce, G. T., Allen, J. W., Doerr, C. L., et al. (1987) Relationships between benzo(a)pyrene-DNA adducts levels and genotoxic effects in mammalian cells. Cancer Res. 47, 3388–3395.PubMedGoogle Scholar
  23. 23.
    Wolterbeek, A. P., Roggeband, R., Steenwinkel, M. J., Rutten, A. A., and Baan, R. A. (1993) Formation and repair of benzo[a]pyrene-DNA adducts in cultured hamster tracheal epithelium determined by 32P-postlabeling analysis and unscheduled DNA synthesis. Carcinogenesis 14, 463–467.PubMedCrossRefGoogle Scholar
  24. 24.
    Shou, M., Harvey, R. G., and Penning, T. M. (1993) Reactivity of benzo[a]pyrene-7,8-dione with DNA. Evidence for the formation of deoxyguanosine adducts. Carcinogenesis 14, 475–482.PubMedCrossRefGoogle Scholar
  25. 25.
    Tang, M. S., Pierce, J. R., Doisy, R. P., Nazimiec, M. E., and Macleod, M. C. (1992) Differences and similarities in the repair of two benzo[a]pyrene diol epoxide isomers induced DNA adducts by uvrA, uvrB, and uvrC gene products. Biochemistry 31, 8429–8436.PubMedCrossRefGoogle Scholar
  26. 26.
    Chatham Workshop Conference (1997) Chatham Workshop Conference on Karyological Monitoring of Normal Cell Populations. International Association of Biological Standardization. Cape Cod, MA.Google Scholar
  27. 27.
    Hsu, T. C, Wu, X., and Trizna, Z. (1996) Mutagen sensitivity in humans—a comparison between two nomenclature systems for recording chromatid breaks. Cancer Genet. Cytogenet. 87, 127–132.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc. 2005

Authors and Affiliations

  • Xifeng Wu
    • 1
  • Yun-Ling Zheng
    • 2
  • T. C. Hsu
    • 3
  1. 1.Department of EpidemiologyM. D. Anderson Cancer CenterHouston
  2. 2.Laboratory of Human CarcinogenesisNational Cancer InstituteBethesda
  3. 3.Department of Cancer BiologyM. D. Anderson Cancer CenterHouston

Personalised recommendations