Comparative aneugenicity of doxorubicin and its derivative idarubicin using fluorescence in situ hybridization techniques

The present study was designed to evaluate and compare the aneugenicity of idarubicin and doxorubicin, topoisomerase-targeting anticancer anthracyclines, using fluorescence in situ hybridization techniques. It was found that idarubicin and doxorubicin treatment (12 mg/kg) induced sperm meiotic delay of 24 h. To determine the frequencies of disomic and diploid sperm, groups of 5 male Swiss albino mice were treated with 3, 6 and 12 mg/kg idarubicin or doxorubicin. Significant increases in the frequencies of disomic and diploid sperm were caused by treatment with all doses of idarubicin and the two highest doses of doxorubicin compared with the controls. Moreover, both compounds significantly increased the frequency of diploid sperm, indicating that complete meiotic arrest occurred. The observation that XX- and YY-sperm significantly prevailed XY-sperm indicates missegregation during the second meiotic division. The results suggest also that earlier prophase stages contribute relatively less to idarubicin and doxorubicin-induced aneuploidy. Effects of the same doses were investigated by the bone-marrow micronucleus test. Significant increases in the frequencies of micronuclei were found after treatment with all doses of both compounds. The responses were also directly correlated with bone marrow suppression. Idarubicin was more toxic than doxorubicin. Exposure to 12 mg/kg of idarubicin and doxorubicin yielded 3.82 and 2.64% micronuclei, respectively, and of these an average of 58.3 and 62.8%, respectively, showed centromeric signals, indicating their formation by whole chromosomes and reflecting the aneugenic activity of both compounds. Correspondingly, about 41.7 and 37.2% of the induced micronuclei, respectively, were centromere-negative, demonstrating that both compounds not only induce chromosome loss but also DNA strand breaks. Based on our data, aneuploidy assays such as sperm-fluorescence in situ hybridization assay and micronucleus test complemented by fluorescence in situ hybridization with centromeric DNA probes have been to some extent validated to be recommended for the assessment of aneuploidogenic effects of chemicals.

► The aneugenicity of idarubicin and doxorubicin was investigated by FISH techniques.
► It was found that both compounds induced meiotic delay of at least 24 h.
► Both compounds increase disomic and diploid sperm and earlier prophase stages contribute relatively less.
► Both compounds are aneugens as well as clastogens in somatic cells as detected by DNA centromeric probe.
► Idarubicin was more toxic than doxorubicin in somatic and germinal cells.