Damage to cellular and isolated DNA induced by a metabolite of aspirin.
Aspirin has been proposed as a possible chemopreventive agent. On the other hand, a recent cohort study showed that aspirin may increase the risk for pancreatic cancer. To clarify whether aspirin is potentially carcinogenic, we investigated the formation of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG), which is correlated with the incidence of cancer, in cultured cells treated with 2,3-dihydroxybenzoic acid (2,3-DHBA), a metabolite of aspirin. 2,3-DHBA induced 8-oxodG formation in the PANC-1 human pancreatic cancer cell line. 2,3-DHBA-induced DNA single-strand breaks were also revealed by comet assay using PANC-1 cells. Flow cytometric analyses showed that 2,3-DHBA increased the levels of intracellular reactive oxygen species (ROS) in PANC-1 cells. The 8-oxodG formation and ROS generation were also observed in the HL-60 leukemia cell line, but not in the hydrogen peroxide (H(2)O(2))-resistant clone HP100 cells, suggesting the involvement of H(2)O(2). In addition, an hprt mutation assay supported the mutagenicity of 2,3-DHBA. We investigated the mechanism underlying the 2,3-DHBA-induced DNA damage using (32)P-labeled DNA fragments of human tumor suppressor genes. 2,3-DHBA induced DNA damage in the presence of Cu(II) and NADH. DNA damage induced by 2,3-DHBA was enhanced by the addition of histone peptide-6 [AKRHRK]. Interestingly, 2,3-DHBA and histone peptide-6 caused base damage in the 5'-ACG-3' and 5'-CCG-3' sequences, hotspots of the p53 gene. Bathocuproine, a Cu(I) chelator, and catalase inhibited the DNA damage. Typical hydroxyl radical scavengers did not inhibit the DNA damage. These results suggest that ROS derived from the reaction of H(2)O(2) with Cu(I) participate in the DNA damage. In conclusion, 2,3-DHBA induces oxidative DNA damage and mutations, which may result in carcinogenesis.
Oikawa
Shinji
S
Department of Environmental and Molecular Medicine, Mie University Graduate School of Medicine, Mie 514-8507, Japan. s-oikawa@doc.medic.mie-u.ac.jp
Kobayashi
Hatasu
H
Tada-Oikawa
Saeko
S
Isono
Yoshiaki
Y
Kawanishi
Shosuke
S
D000080242:8-Hydroxy-2'-Deoxyguanosine; D001241:Aspirin / Q000378:metabolism / Q000633:toxicity*; D045744:Cell Line, Tumor; D020552:Comet Assay; D003300:Copper / Q000633:toxicity; D053904:DNA Breaks, Single-Stranded / Q000187:drug effects; D004249:DNA Damage; D004273:DNA, Neoplasm / Q000187:drug effects / Q000235:genetics; D003849:Deoxyguanosine / Q000031:analogs & derivatives / Q000378:metabolism; D016147:Genes, Tumor Suppressor; D018922:HL-60 Cells; D006801:Humans; D062385:Hydroxybenzoates / Q000378:metabolism / Q000633:toxicity; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics; D066298:In Vitro Techniques; D008954:Models, Biological; D009243:NAD / Q000494:pharmacology; D010190:Pancreatic Neoplasms / Q000209:etiology / Q000235:genetics / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism
D004273:DNA, Neoplasm; D062385:Hydroxybenzoates; D017382:Reactive Oxygen Species; D009243:NAD; C009135:2,3-dihydroxybenzoic acid; D003300:Copper; D000080242:8-Hydroxy-2'-Deoxyguanosine; D007041:Hypoxanthine Phosphoribosyltransferase; D003849:Deoxyguanosine; D001241:Aspirin