Formation and stability of alkylated pyrimidines and purines (including imidazole ring-opened 7-alkylguanine) and alkylphosphotriesters in liver DNA of adult rats treated with ethylnitrosourea or dimethylnitrosamine. 1986

L Den Engelse, and G J Menkveld, and R J De Brij, and A D Tates

Adult rats received a single injection of the carcinogens N-ethyl-N-nitrosourea (ENU) or N,N-dimethylnitrosamine (DMN) and were killed after various time intervals (up to 56 days). Liver DNA was analysed by h.p.l.c. for alkylated purines [3-alkyladenine (3-alkylAde), 7-alkylAde, 3-alkylguanine (3-alkylGua), O6-alkylGua, 7-alkylGua, imidazole ring-opened (ro) 7-alkylGua] and alkylated pyrimidines [O2-alkylcytosine (O2-alkylCyt), 3-alkylCyt, O2-alkylthymine (O2-alkylThy), 3-alkylThy, O4-alkylThy]. Alkylphosphotriesters were assayed by both h.p.l.c. [alkylphosphotriester of thymidyl(3'-5')thymidine, (dTp(alkyl)dT)] and alkaline sucrose sedimentation (total phosphotriesters). No significant amounts of 3-alkylCyt and ro 7-alkylGua were observed, but the presence of all other products could be established 2 h after DMN or ENU. At that moment the relative amounts of some products (O2-alkylCyt, O2-alkylThy, O4-alkylThy, O6-alkylGua), when compared with 7-alkylGua, were smaller than those observed after in vitro alkylation of isolated DNA. This suggests either a lower accessibility of some sites in DNA in situ, or (as known already for O6-alkylGua) the presence of rapidly exhausted, fast repair modes for these adducts. The in vivo stability of ethylated products was (much) higher than that of the homologous methylated products. In the case of O-alkylated thymidines this difference was impressive: apparent half-life values of 17 days for O2-ethylthymine (O2-EtThy) and 14 days for O4-EtThy were calculated, whereas the corresponding values for O2-methylthymine (O2-MeThy) and O4-MeThy were 12 h and less than 4 h, respectively. Substantial, although smaller differences were also found for O2-alkylCyt (86 versus less than 4 h), 3-alkylGua (104 versus 17 h) and dTp(alkyl)dT (32 versus 7 days), whereas the rates of removal of 7-EtGua and 7-MeGua differed by 2.5-5 times (depending on the period compared). At 56 days after ENU the two major lesions were the ethylphosphotriester and O2-EtThy, whereas only traces of 7-EtGua were observed. Parenchymal and non-parenchymal liver cells of some DMN-treated rats were separated before DNA isolation. It was found that the extents of DNA alkylation at 6 h after DMN administration were almost identical, indicating that DMN activation is the same for both cell types. The results are discussed in relation to the carcinogenic effects of methylating and ethylating agents.

UI MeSH Term Description Entries
D007700 Kinetics The rate dynamics in chemical or physical systems.
D008099 Liver A large lobed glandular organ in the abdomen of vertebrates that is responsible for detoxification, metabolism, synthesis and storage of various substances. Livers
D008297 Male Males
D008745 Methylation Addition of methyl groups. In histo-chemistry methylation is used to esterify carboxyl groups and remove sulfate groups by treating tissue sections with hot methanol in the presence of hydrochloric acid. (From Stedman, 25th ed) Methylations
D008770 Methylnitrosourea A nitrosourea compound with alkylating, carcinogenic, and mutagenic properties. Nitrosomethylurea,N-Methyl-N-nitrosourea,NSC-23909,N Methyl N nitrosourea,NSC 23909,NSC23909
D010710 Phosphates Inorganic salts of phosphoric acid. Inorganic Phosphate,Phosphates, Inorganic,Inorganic Phosphates,Orthophosphate,Phosphate,Phosphate, Inorganic
D011687 Purines A series of heterocyclic compounds that are variously substituted in nature and are known also as purine bases. They include ADENINE and GUANINE, constituents of nucleic acids, as well as many alkaloids such as CAFFEINE and THEOPHYLLINE. Uric acid is the metabolic end product of purine metabolism.
D011743 Pyrimidines A family of 6-membered heterocyclic compounds occurring in nature in a wide variety of forms. They include several nucleic acid constituents (CYTOSINE; THYMINE; and URACIL) and form the basic structure of the barbiturates.
D002851 Chromatography, High Pressure Liquid Liquid chromatographic techniques which feature high inlet pressures, high sensitivity, and high speed. Chromatography, High Performance Liquid,Chromatography, High Speed Liquid,Chromatography, Liquid, High Pressure,HPLC,High Performance Liquid Chromatography,High-Performance Liquid Chromatography,UPLC,Ultra Performance Liquid Chromatography,Chromatography, High-Performance Liquid,High-Performance Liquid Chromatographies,Liquid Chromatography, High-Performance
D004128 Dimethylnitrosamine A nitrosamine derivative with alkylating, carcinogenic, and mutagenic properties. It causes serious liver damage and is a hepatocarcinogen in rodents. Nitrosodimethylamine,N-Nitrosodimethylamine,NDMA Nitrosodimethylamine,N Nitrosodimethylamine,Nitrosodimethylamine, NDMA

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