Methoxyamine modification of abasic sites protects CHO cells from the cytotoxic and mutagenic effects of oxygen alkylation.
The biological effects of the interaction of methoxyamine (MX) with apurinic/apyrimidinic (AP) sites produced in CHO cells by treatment with alkylating agents were examined. A decrease in cytotoxicity was observed after a 10 min treatment with the SN1 alkylating agents ethylnitrosourea (ENU), N-ethyl-N'-nitro-N-nitrosoguanidine (ENNG) and N-methyl-nitrosourea when MX was present in the culture medium. Furthermore MX reduced the number of mutations to 6-thioguanine resistance induced by ENU and ENNG and the number of sister chromatid exchanges induced by ENU. In contrast, no protective effect of MX on survival was observed after a 10 min treatment with the SN2 alkylating agents diethylsulfate (DES), ethyl methane sulfonate and methyl methane sulfonate. A 3 h exposure to MX abolished the protective effect of MX on ENU-induced cytotoxicity and increased the cytotoxicity of DES. In vitro studies with synthetic oligonucleotides containing a single AP site opposite a normal guanine or O6-methylguanine showed that MX inhibits the cleavage of AP sites by the CHO AP endonuclease(s). A model is proposed in which different DNA lesions are involved in AP site formation after treatment with SN2 or SN2 alkylating agents. The involvement of specific alkylation products in cytotoxicity and mutagenesis is also discussed.
Fortini
P
P
Laboratory of Comparative Toxicology and Ecotoxicology, Istituto Superiore di Sanita, Rome, Italy.
Rosa
S
S
Zijno
A
A
Calcagnile
A
A
Bignami
M
M
Dogliotti
E
E
D000477:Alkylating Agents / Q000633:toxicity*; D000818:Animals; D001483:Base Sequence; D016466:CHO Cells / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D006224:Cricetinae; D004247:DNA / Q000378:metabolism*; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase; D043223:Deoxyribonuclease IV (Phage T4-Induced); D004706:Endodeoxyribonucleases / Q000494:pharmacology; D006898:Hydroxylamines / Q000494:pharmacology*; D008969:Molecular Sequence Data; D009154:Mutation; D009841:Oligonucleotides / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism*; D012854:Sister Chromatid Exchange / Q000187:drug effects
D000477:Alkylating Agents; D006898:Hydroxylamines; D009841:Oligonucleotides; D004247:DNA; C005214:methoxyamine; D004706:Endodeoxyribonucleases; D043223:Deoxyribonuclease IV (Phage T4-Induced); D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase; D010100:Oxygen