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    Parallel evaluation of Doxorubicin inducing Genetic damage in human lymphocytes and sperm using the Comet assay and spectral karyotyping

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    Publication date
    2004
    Author
    Anderson, Diana
    Baumgartner, Adolf
    Cemeli, Eduardo
    Schmid, Thomas E.
    Keyword
    Genetic damage
    Fish method
    DNA damage
    Human sperm
    Lymphocytes
    Chromosome aberrations
    Comet assay
    Sky analysis
    Peer-Reviewed
    Yes
    
    Metadata
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    Abstract
    In recent years, two techniques for detecting genetic damage in the whole genome have gained importance: the alkaline comet assay, to detect DNA damage such as strand breaks and alkali-labile sites, and a multicolour FISH method, spectral karyotyping (SKY), to identify chromosomal aberrations simultaneously in all metaphase chromosomes. In the present study, the induction of DNA damage in human sperm and lymphocytes in vitro has been studied employing an anticancer drug, doxorubicin (DX). An increase in DNA damage was observed with the comet assay as the median per cent head DNA of sperm significantly decreased from 82.07 and 85.14% in the untreated control groups to 63.48 and 72.52% at doses of 0.8 µM DX. At 1.6 µM the percentage declined to 60.96% (the corresponding tail moment increased from 4.42 to 12.19). In stimulated lymphocytes, a significant increase was observed in tail moment, from 0.72 and 0.53 in controls to 15.17 and 12.10 at 0.2 µM DX, continuing at the same level to a final concentration of 1.6 µM. Structural aberrations found in the parallel SKY study in stimulated lymphocytes at 0.2 µM DX consisted of 14% chromatid-type and 2% chromosome-type aberrations; none were found in controls. The SKY results correlate very well with the findings of the comet assay in lymphocytes where DNA damage was observed at similar doses. This study is the first reporting use of the comet assay and SKY analysis in parallel after chemical treatment. The potential of the two techniques together is evident, as they represent a set of assays feasible for evaluating damage in human somatic and germ cells after chemical treatment (i) by direct observation of two different end-points, detecting general DNA damage and chromosomal aberrations and (ii) by extrapolation from lymphocytes to sperm, which provides a `parallelogram¿ approach in human cells.
    URI
    http://hdl.handle.net/10454/2700
    Version
    No full-text available in the repository
    Citation
    Anderson, D., Baumgartner, A., Cemeli, E. and Schmid, T.E. (2004). Parallel evaluation of Doxorubicin inducing Genetic damage in human lymphocytes and sperm using the Comet assay and spectral karyotyping. Mutagenesis. Vol.19, No.4, pp.313-318.
    Link to publisher’s version
    http://mutage.oxfordjournals.org/cgi/content/full/19/4/313
    Type
    Article
    Collections
    Life Sciences Publications

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