title: Effects of ethylene oxide and ethylene inhalation on DNA adducts, apurinic/apyrimidinic sites and expression of base excision DNA repair genes in rat brain, spleen, and liver.
abstract: Ethylene oxide (EO) is an important industrial chemical that is classified as a known human carcinogen (IARC, Group 1). It is also a metabolite of ethylene (ET), a compound that is ubiquitous in the environment and is the most used petrochemical. ET has not produced evidence of cancer in laboratory animals and is "not classifiable as to its carcinogenicity to humans" (IARC, Group 3). The mechanism of carcinogenicity of EO is not well characterized, but is thought to involve the formation of DNA adducts. EO is mutagenic in a variety of in vitro and in vivo systems, whereas ET is not. Apurinic/apyrimidinic sites (AP) that result from chemical or glycosylase-mediated depurination of EO-induced DNA adducts could be an additional mechanism leading to mutations and chromosomal aberrations. This study tested the hypothesis that EO exposure results in the accumulation of AP sites and induces changes in expression of genes for base excision DNA repair (BER). Male Fisher 344 rats were exposed to EO (100 ppm) or ET (40 or 3000 ppm) by inhalation for 1, 3 or 20 days (6h/day, 5 days a week). Animals were sacrificed 2h after exposure for 1, 3 or 20 days as well as 6, 24 and 72 h after a single-day exposure. Experiments were performed with tissues from brain and spleen, target sites for EO-induced carcinogenesis, and liver, a non-target organ. Exposure to EO resulted in time-dependent increases in N7-(2-hydroxyethyl)guanine (7-HEG) in brain, spleen, and liver and N7-(2-hydroxyethyl)valine (7-HEVal) in globin. Ethylene exposure also induced 7-HEG and 7-HEVal, but the numbers of adducts were much lower. No increase in the number of aldehydic DNA lesions, an indicator of AP sites, was detected in any of the tissues between controls and EO-, or ET-exposed animals, regardless of the duration or strength of exposure. EO exposure led to a 3-7-fold decrease in expression of 3-methyladenine-DNA glycosylase (Mpg) in brain and spleen in rats exposed to EO for 1 day. Expression of 8-oxoguanine DNA glycosylase, Mpg, AP endonuclease (Ape), polymerase beta (Pol beta) and alkylguanine methyltransferase were increased by 20-100% in livers of rats exposed to EO for 20 days. The only effects of ET on BER gene expression were observed in brain, where Ape and Pol beta expression were increased by less than 20% after 20 days of exposure to 3000 ppm. These data suggest that DNA damage induced by exposure to EO is repaired without accumulation of AP sites and is associated with biologically insignificant changes in BER gene expression in target organs. We conclude that accumulation of AP sites is not a likely primary mechanism for mutagenicity and carcinogenicity of EO.
authors.affiliation: Department of Environmental Sciences and Engineering, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
authors.forename: Ivan
authors.initials: I
authors.lastname: Rusyn
authors.forename: Shoji
authors.initials: S
authors.lastname: Asakura
authors.forename: Yutai
authors.initials: Y
authors.lastname: Li
authors.forename: Oksana
authors.initials: O
authors.lastname: Kosyk
authors.forename: Hasan
authors.initials: H
authors.lastname: Koc
authors.forename: Jun
authors.initials: J
authors.lastname: Nakamura
authors.forename: Patricia B
authors.initials: PB
authors.lastname: Upton
authors.forename: James A
authors.initials: JA
authors.lastname: Swenberg
mesh_terms: D000280:Administration, Inhalation; D000818:Animals; D017209:Apoptosis / Q000235:genetics; D001921:Brain / Q000378:metabolism; D001923:Brain Chemistry; D004247:DNA / Q000187:drug effects; D018736:DNA Adducts / Q000032:analysis / Q000378:metabolism*; D004260:DNA Repair / Q000235:genetics*; D005027:Ethylene Oxide / Q000008:administration & dosage / Q000633:toxicity*; D005030:Ethylenes / Q000008:administration & dosage / Q000633:toxicity*; D015870:Gene Expression / Q000187:drug effects*; D006147:Guanine / Q000031:analogs & derivatives / Q000032:analysis / Q000378:metabolism; D008099:Liver / Q000737:chemistry / Q000378:metabolism; D008297:Male; D051381:Rats; D011916:Rats, Inbred F344; D013154:Spleen / Q000737:chemistry / Q000378:metabolism
chemical_list: D018736:DNA Adducts; D005030:Ethylenes; C035018:N(7)-hydroxyethylguanine; D006147:Guanine; D004247:DNA; C036216:ethylene; D005027:Ethylene Oxide