title: Induction of DNA strand breaks by trihalomethanes in primary human lung epithelial cells.
abstract: Trihalomethanes (THMs) are disinfection by-products and suspected human carcinogens present in chlorinated drinking water. Previous studies have shown that many THMs induce sister chromatid exchanges and DNA strand breaks in human peripheral blood lymphocytes in vitro. Exposure to THMs occurs through oral, dermal, or inhalation routes, with the lung being a target of exposure by the latter route, although not a target for rodent carcinogenicity. Thus, to examine the genotoxicity of THMs in this tissue, we used the comet assay to examine the DNA damaging ability of five THMs in primary human lung epithelial cells. Cells were collected by scraping the large airways of four volunteers with a cytology brush and then passaging the cells no more than three times in order to have sufficient numbers for the experiments. Cells were exposed for 3h to 10, 100, or 1000 microM CHCl(3), CHCl(2)Br, CHClBr(2), or CHBr(3); CH(2)Cl(2) was also used as a model dihalomethane for comparison to the THMs. The compounds ranked as follows for DNA damaging ability: CHCl(2)Br>CHBr(3)>CHCl(3) approximately equal CH(2)Cl(2); CHClBr(2) was negative. Considerable inter-individual variation was observed. For example, CHCl(3) was genotoxic in only two subjects, and the interaction between dose and donor was highly significant (P<0.001). The same variation was observed for CHCl(2)Br, which was positive only in the two subjects in which CHCl(3) was negative. This variation was not due to the GSTT1-1 genotype of the subjects. Although two subjects were GSTT1-1(+), and two were GSTT1-1(-), no cultured cells with a GSTT1-1(+) genotype had detectable GSTT1-1 enzymatic activity nor did any frozen epithelial cells that had not been cultured. However, GSTT1-1 enzymatic activity was detected in fresh (neither frozen nor cultured) lung cells. These results show that freezing or culturing causes lung cells to lose GSTT1-1 activity and that factors other than GSTT1-1 activity play a role in the variable responses of these human cells to the genotoxicity of the halomethanes studied here.
authors.affiliation: US Environmental Protection Agency, National Health and Environmental, Effects Research Laboratory, Research Triangle Park, NC 27711, USA.
authors.forename: Stefano
authors.initials: S
authors.lastname: Landi
authors.forename: Alessio
authors.initials: A
authors.lastname: Naccarati
authors.forename: Matthew K
authors.initials: MK
authors.lastname: Ross
authors.forename: Nancy M
authors.initials: NM
authors.lastname: Hanley
authors.forename: Lisa
authors.initials: L
authors.lastname: Dailey
authors.forename: Robert B
authors.initials: RB
authors.lastname: Devlin
authors.forename: Marie
authors.initials: M
authors.lastname: Vasquez
authors.forename: Rex A
authors.initials: RA
authors.lastname: Pegram
authors.forename: David M
authors.initials: DM
authors.lastname: DeMarini
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D020552:Comet Assay; D003600:Cytosol / Q000187:drug effects / Q000201:enzymology; D004249:DNA Damage; D004305:Dose-Response Relationship, Drug; D020022:Genetic Predisposition to Disease; D005838:Genotype; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D008099:Liver / Q000187:drug effects / Q000201:enzymology; D051379:Mice; D009153:Mutagens / Q000633:toxicity*; D011817:Rabbits; D020545:Respiratory Mucosa / Q000187:drug effects* / Q000201:enzymology; D022882:Trihalomethanes / Q000633:toxicity*
chemical_list: D009153:Mutagens; D022882:Trihalomethanes; C413545:glutathione S-transferase T1; D005982:Glutathione Transferase