Cytogenetic damage induced by acrylamide and glycidamide in mammalian cells: correlation with specific glycidamide-DNA adducts.
Acrylamide (AA) is a suspected human carcinogen generated in carbohydrate-rich foodstuffs upon heating. Glycidamide (GA), formed via epoxidation, presumably mediated by cytochrome P450 2E1, is thought to be the active metabolite playing a central role in AA genotoxicity. In this work we investigated DNA damage induced by AA and GA in mammalian cells, using V79 Chinese hamster cells. For this purpose, we evaluated two cytogenetic end points, chromosomal aberrations (CAs) and sister chromatid exchanges (SCEs), as well as the levels of specific GA-DNA adducts, namely, N7-(2-carbamoyl-2-hydroxyethyl)guanine (N7-GA-Gua) and N3-(2-carbamoyl-2-hydroxyethyl)adenine (N3-GA-Ade) using high-performance liquid chromatography coupled with tandem mass spectrometry. GA was more cytotoxic and clastogenic than AA. Both AA and GA induced CAs (breaks and gaps) and decreased the mitotic index. GA induced SCEs in a dose-responsive manner; with AA, SCEs were increased at only the highest dose tested (2mM). A linear dose-response relationship was observed between the GA concentration and the levels of N7-GA-Gua. This adduct was detected for concentrations as low as 1 microM GA. N3-GA-Ade was also detected, but only at very high GA concentrations (>or= 250 microM). There was a very strong correlation between the levels of N7-GA-Gua in the GA- and AA-treated cells and the extent of SCE induction. Such correlation was not apparent for CAs. These data suggest that the induction of SCEs by AA is associated with the metabolism of AA to GA and subsequent formation of depurinating DNA adducts; however, other mechanisms must be involved in the induction of CAs.
Martins
Célia
C
Department of Genetics, Faculty of Medical Sciences, New University of Lisbon, 1349-008 Lisboa, Portugal.
Oliveira
Nuno G
NG
Pingarilho
Marta
M
Gamboa da Costa
Gonçalo
G
Martins
Vanda
V
Marques
M Matilde
MM
Beland
Frederick A
FA
Churchwell
Mona I
MI
Doerge
Daniel R
DR
Rueff
José
J
Gaspar
Jorge Francisco
JF
D020106:Acrylamide / Q000633:toxicity*; D000225:Adenine / Q000031:analogs & derivatives* / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D002869:Chromosome Aberrations / Q000139:chemically induced*; D006224:Cricetinae; D003412:Cricetulus; D018736:DNA Adducts / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004852:Epoxy Compounds / Q000633:toxicity*; D006147:Guanine / Q000031:analogs & derivatives* / Q000378:metabolism; D015394:Molecular Structure; D009153:Mutagens / Q000633:toxicity*; D012854:Sister Chromatid Exchange / Q000187:drug effects*
D018736:DNA Adducts; D004852:Epoxy Compounds; D009153:Mutagens; C518583:N3-(2-carbamoyl-2-hydroxyethyl)adenine; D020106:Acrylamide; C089230:N7-(2-((hydroxyethyl)thio)ethyl)guanine; D006147:Guanine; C071834:glycidamide; D000225:Adenine