Lack of direct involvement of 8-hydroxy-2'-deoxyguanosine in hypoxanthine-guanine phosphoribosyltransferase mutagenesis in V79 cells treated with N,N'-bis(2-hydroxyperoxy-2-methoxyethyl)-1,4,5,8-naphthalenetetracarboxylic-diimide (NP-III) or riboflavin.
The object of this study is to investigate the relationship between a typical product of oxidative DNA damage, 8-hydroxy-2'-deoxyguanosine (8OHdG), and mutagenesis in V79 cells through a molecular analysis of hypoxanthine-guanine phosphoribosyltransferase (hprt) gene mutants. We performed a direct sequencing analysis of the cDNA of mutants obtained after treatment with N,N'-bis(2-hydroxyperoxy-2-methoxyethyl)-1,4,5,8-naphthalenetetracarboxylic-diimide (NP-III) or riboflavin, each of which induces the formation of 8OHdG in cellular DNA upon UVA irradiation. The frequency of mutation after both treatments was no more than 2 to 5 times the control value. A considerable number of the mutants could not be amplified by RT-PCR, and this was also the case for the control mutants. Among the mutants analyzed, deletions and a TA-->AT transversion occurred predominantly. The reasons for the weak association of induction of 8OHdG with frequency of mutation and the possible mechanism of oxidative-stress-derived mutagenesis are discussed.
Nakajima
Madoka
M
Department of Environmental and Preventive Medicine, Kanazawa University Graduate School of Medical Science, Kanazawa, Ishikawa 920-8640, Japan.
Takeuchi
Toru
T
Ogino
Keiki
K
Morimoto
Kanehisa
K
D000080242:8-Hydroxy-2'-Deoxyguanosine; D000818:Animals; D002478:Cells, Cultured / Q000187:drug effects / Q000528:radiation effects; D006224:Cricetinae; D004247:DNA / Q000528:radiation effects; D004249:DNA Damage; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000378:metabolism*; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics* / Q000378:metabolism; D008168:Lung / Q000187:drug effects* / Q000378:metabolism; D016296:Mutagenesis; D009281:Naphthalenes / Q000494:pharmacology*; D018384:Oxidative Stress; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012256:Riboflavin / Q000494:pharmacology*
D009281:Naphthalenes; C067580:N,N'-bis(2-hydroxyperoxy-2-methoxyethyl)-1,4,5,8-naphthalenetetracarboxylic diimide; D000080242:8-Hydroxy-2'-Deoxyguanosine; D004247:DNA; D007041:Hypoxanthine Phosphoribosyltransferase; D003849:Deoxyguanosine; D012256:Riboflavin