A quinoxaline 1,4-di- N-oxide derivative induces DNA oxidative damage not attenuated by vitamin C and E treatment

• Published in: Chemico-biological interactions 2007-06, Vol.168 (2), p.95-105
• Main Authors: Azqueta, Amaia, Arbillaga, Leire, Pachón, Gisela, Cascante, Marta, Creppy, Edmond E, Cerain, Adela López de
• Format: Article
• Language: English
• Subjects: Ascorbic Acid - pharmacology; Bioreductive agent; Caco-2 Cells; Comet assay; DNA Damage; DNA Glycosylases - metabolism; DNA oxidative damage; DNA-Formamidopyrimidine Glycosylase - metabolism; Humans; Hypoxia - metabolism; Prodrugs - pharmacology; Quinoxalines; Quinoxalines - pharmacology; Reactive Oxygen Species - metabolism; Vitamin C; Vitamin E; Vitamin E - pharmacology
• ISSN: 0009-2797; 1872-7786
• DOI: 10.1016/j.cbi.2007.02.013

Some anticancer compounds are pro-drugs which give rise to toxic species through enzymatic reduction. The quinoxaline-di- N-oxide derivative Q-85 HCl (7-chloro-3-[[( N, N-dimethylamino)propyl]amino]-2-quinoxalinecarbonitrile 1,4-di- N-oxide hydrochloride) is a bioreductive compound selectively toxic in hypoxia. Due to the possibility of secondary tumors the study of the genotoxic capability of antitumoral drugs is very important. The aim of this study was to assess the ability of Q-85 HCl to produce reactive oxygen species (ROS) and oxidative DNA damage in Caco-2 cells, both in hypoxia and in well-oxygenated conditions. Secondly, we attempted to evaluate the effect of vitamins C and E under hypoxic and normoxic conditions, in order to determine if these antioxidant substances modify Q-85 HCl effect in hypoxic cells or possibly exert a protective action in normal cells. Caco-2 cells were treated with Q-85 HCl for 2 h, at high concentrations in normoxia (0.1–5 μM) and at low concentrations in hypoxia (0.002–0.1 μM). In normoxia, a dose-related significant increase in intracellular ROS level was evident; in hypoxia all the concentrations produced very high level of ROS. Just after the treatment and 24 h later, oxidative DNA damage was evaluated by the modified comet assay after post-digestion of the cells with formamidopyrimidine-DNA glycosylase (FPG) and endonuclease III (Endo III). Q-85 HCl treatment evoked a significant dose-dependent increase in the total comet score of the cells both in hypoxia and normoxia, indicating that this compound or some metabolite is able to oxidize purine and pyrimidine bases. After 24 h DNA damage caused by the compound was completely repaired with only one exception: cells treated with the highest concentration of Q-85 HCl in hypoxia and post-digested with FPG. Vitamin C (5–100 μM) and vitamin E (500–400 μM) did not have a pro-oxidant effect in Caco-2 cells. Treatment of cells with vitamin C (10 μM) or vitamin E (100 μM) did not significantly reduce oxidative DNA damage in hypoxia and normoxia. In conclusion, the use of these vitamins would not hinder toxicity against hypoxic cells, but a protective effect in normoxic cells was not evident.