title: lacZ mouse embryonic fibroblasts detect both clastogens and mutagens.
abstract: The clastogenic effects of MMC and BLM and the mutagenic effects of B[a]P, N-ac-AAF and ENU were studied in mouse embryonic fibroblasts derived from wild-type (WT) and Rad54/Rad54B-deficient mice. Clastogens as well as mutagens showed a statistically significant induction of mutations in the lacZ reporter gene both in a WT and Rad54/Rad54B-deficient genetic background. Rad54/Rad54B MEFs appeared equally sensitive to the clastogens compared to WT MEFs, except for MMC. The type of mutations induced by the different compounds was investigated further by hybridizing the mutant colonies with total mouse DNA. An obvious increased number of mouse DNA positive clones was observed after BLM and MMC exposure, indicating that after these treatments genome rearrangements/translocations had occurred. In this hybridization assay, Rad54/Rad54B MEFs did not show more rearrangements/translocations than WT MEFs. As expected, the mutagens used showed no increase in chromosomal rearrangements or transloctions in MEFs derived from both genotypes. These results show that WT MEFs carrying the lacZ reporter gene on a plasmid are capable to detect both clastogenic as well as mutagenic effects of compounds in vitro. Deletion of the Rad54 and Rad54B genes did not further enhance the sensitivity of MEFs towards clastogens.
authors.affiliation: National Institute of Public Health and the Environment (RIVM), Laboratory for Health Protection Research, Antonie van Leeuwenhoeklaan 9, 3721 MA Bilthoven, The Netherlands.
authors.forename: Anuska G
authors.initials: AG
authors.lastname: Mahabir
authors.forename: Edwin
authors.initials: E
authors.lastname: Zwart
authors.forename: Mirjam
authors.initials: M
authors.lastname: Schaap
authors.forename: Jan
authors.initials: J
authors.lastname: van Benthem
authors.forename: Annemieke
authors.initials: A
authors.lastname: de Vries
authors.forename: Lya G
authors.initials: LG
authors.lastname: Hernandez
authors.forename: Coenraad F M
authors.initials: CF
authors.lastname: Hendriksen
authors.forename: Harry
authors.initials: H
authors.lastname: van Steeg
mesh_terms: D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002869:Chromosome Aberrations; D004249:DNA Damage; D004265:DNA Helicases; D004260:DNA Repair; D005347:Fibroblasts / Q000187:drug effects; D017930:Genes, Reporter; D007763:Lac Operon; D051379:Mice; D008822:Mice, Transgenic; D015162:Micronucleus Tests; D009152:Mutagenicity Tests / Q000379:methods*; D009153:Mutagens; D009154:Mutation; D009687:Nuclear Proteins / Q000235:genetics
chemical_list: D009153:Mutagens; D009687:Nuclear Proteins; D004265:DNA Helicases; C103534:Rad54l protein, mouse