title: Assays to predict the genotoxicity of the chromosomal mutagen etoposide -- focussing on the best assay.
abstract: The topoisomerase II inhibitor etoposide is used routinely to treat a variety of cancers in patients of all ages. As a result of its extensive use in the clinic and its association with secondary malignancies it has become a compound of great interest with regard to its genotoxic activity in vivo. This paper describes a series of assays that were employed to determine the in vivo genotoxicity of etoposide in a murine model system. The alkaline comet assay detected DNA damage in the bone marrow mononuclear compartment over the dose range of 10--100mg/kg and was associated with a large and dose dependent rise in the proportion of cells with severely damaged DNA. In contrast, the bone marrow micronucleus assay was found to be sensitive to genotoxic damage between the doses of 0.1--1mg/kg without any corresponding increases in cytotoxicity. An increase in the mutant frequency was undetectable at the Hprt locus at administered doses of 1 and 10mg/kg of etoposide, however, an increase in the mutant frequency was seen at the Aprt locus at these doses. We conclude that the BMMN assay is a good short-term predictor of the clastogenicity of etoposide at doses that do not result in cytotoxic activity, giving an indication of potential mutagenic effects. Moreover, the detection of mutants at the Aprt locus gives an indication of the potential of etoposide to cause chromosomal mutations that may lead to secondary malignancy.
authors.affiliation: Gene Therapy Group, Christie Hospital (NHS) Trust, Wilmslow Road, Manchester M20 4BX, UK. suzanne.turner@bbsrc.ac.uk
authors.forename: S D
authors.initials: SD
authors.lastname: Turner
authors.forename: S W
authors.initials: SW
authors.lastname: Wijnhoven
authors.forename: H
authors.initials: H
authors.lastname: Tinwell
authors.forename: L S
authors.initials: LS
authors.lastname: Lashford
authors.forename: J A
authors.initials: JA
authors.lastname: Rafferty
authors.forename: J
authors.initials: J
authors.lastname: Ashby
authors.forename: H
authors.initials: H
authors.lastname: Vrieling
authors.forename: L J
authors.initials: LJ
authors.lastname: Fairbairn
mesh_terms: D000228:Adenine Phosphoribosyltransferase / Q000235:genetics; D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000633:toxicity; D020552:Comet Assay; D005047:Etoposide / Q000633:toxicity*; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D051379:Mice; D015162:Micronucleus Tests; D009152:Mutagenicity Tests / Q000379:methods*; D009153:Mutagens / Q000633:toxicity*; D013154:Spleen / Q000187:drug effects / Q000201:enzymology; D059005:Topoisomerase II Inhibitors
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D009153:Mutagens; D059005:Topoisomerase II Inhibitors; D005047:Etoposide; D000228:Adenine Phosphoribosyltransferase; D007041:Hypoxanthine Phosphoribosyltransferase