title: Induction of TK mutations in human lymphoblastoid TK6 cells by the rat carcinogen 3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone (MX).
abstract: 3-Chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone (MX), a chlorine disinfection by-product in drinking water, is carcinogenic in rats and genotoxic in mammalian cells in vitro. In the current study, the mechanism of genotoxicity of MX in human lymphoblastoid TK6 cells was investigated by use of the Comet assay, the micronucleus test, and the thymidine kinase (TK) gene-mutation assay. MX induced a concentration-dependent increase in micronuclei and TK mutations. The lowest effective concentrations in the MN test and the TK gene-mutation assay were 37.5μM and 25μM, respectively. In the Comet assay, a slight although not statistically significant increase was observed in the level of DNA damage induced by MX in the concentration range of 25-62.5μM. Molecular analysis of the TK mutants revealed that MX induced primarily point mutations or other small intragenic mutations (61%), while most of the remaining TK mutants (32%) were large deletions at the TK locus, leading to the hemizygous-type loss-of-heterozygosity (LOH) mutations. These findings show that aside from inducing point mutations, MX also generates LOH at the TK locus in human cells and may thus cause the inactivation of tumour-suppressor genes by LOH.
authors.affiliation: Division of Genetics and Mutagenesis, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan. pasi.hakulinen@thi.fi
authors.forename: Pasi
authors.initials: P
authors.lastname: Hakulinen
authors.forename: Ayumi
authors.initials: A
authors.lastname: Yamamoto
authors.forename: Naoki
authors.initials: N
authors.lastname: Koyama
authors.forename: Wakako
authors.initials: W
authors.lastname: Kumita
authors.forename: Manabu
authors.initials: M
authors.lastname: Yasui
authors.forename: Masamitsu
authors.initials: M
authors.lastname: Honma
mesh_terms: D000818:Animals; D002273:Carcinogens / Q000633:toxicity*; D002460:Cell Line; D005663:Furans / Q000633:toxicity*; D006801:Humans; D019656:Loss of Heterozygosity; D009152:Mutagenicity Tests; D009153:Mutagens / Q000633:toxicity*; D009154:Mutation; D051381:Rats; D013937:Thymidine Kinase / Q000235:genetics*
chemical_list: D002273:Carcinogens; D005663:Furans; D009153:Mutagens; C054121:3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone; D013937:Thymidine Kinase