Department of Biochemistry and Molecular Biology, and ‡Department of Pharmacology, College of Medicine, Pennsylvania State University , Hershey, Pennsylvania 17033, United States.
Chem Res Toxicol. 2013 Nov 18;26(11):1746-54. doi: 10.1021/tx400296x. Epub 2013 Oct 30.
Previous studies in rats, mice, and in vitro systems showed that 6-NC can be metabolically activated by two major pathways: (1) the formation of N-hydroxy-6-aminochrysene by nitroreduction to yield three major adducts, N-(dG-8-yl)-6-AC, 5-(dG-N(2)-yl)-6-AC, and N-(dA-8-yl)-6-AC, and (2) the formation of trans-1,2-dihydroxy-1,2-dihydro-6-hydroxylaminochrysene (1,2-DHD-6-NHOH-C) by a combination of nitroreduction and ring oxidation pathways to yield N-(dG-8-yl)-1,2-DHD-6-AC, 5-(dG-N(2)-yl)-1,2-DHD-6-AC and N-(dA-8-yl)-1,2-DHD-6-AC. These DNA lesions are likely to cause mutations if they are not removed by cellular defense mechanisms before DNA replication occurs. Here, we compared for the first time, in HeLa cell extracts in vitro, the relative nucleotide excision repair (NER) efficiencies of DNA lesions derived from simple nitroreduction and from a combination of nitroreduction and ring oxidation pathways. We show that the N-(dG-8-yl)-1,2-DHD-6-AC adduct is more resistant to NER than the N-(dG-8-yl)-6-AC adduct by a factor of ∼2. Furthermore, the N-(dA-8-yl)-6-AC is much more resistant to repair since its NER efficiency is ∼8-fold lower than that of the N-(dG-8-yl)-6-AC adduct. On the basis of our previous study and the present investigation, lesions derived from 6-NC and benzo[a]pyrene can be ranked from the most to the least resistant lesion as follows: N-(dA-8-yl)-6-AC > N-(dG-8-yl)-1,2-DHD-6-AC > 5-(dG-N(2)-yl)-6-AC ≃ N-(dG-8-yl)-6-AC ≃ (+)-7R,8S,9S,10S-benzo[a]pyrene diol epoxide-derived trans-anti-benzo[a]pyrene-N(2)-dG adduct. The slow repair of the various lesions derived from 6-NC and thus their potential persistence in mammalian tissue could in part account for the powerful carcinogenicity of 6-NC as compared to B[a]P in the rat mammary gland.
先前在大鼠、小鼠和体外系统中的研究表明,6-NC 可以通过两种主要途径进行代谢激活:(1)通过硝基还原形成 N-羟基-6-氨基苊,生成三种主要加合物,N-(dG-8-基)-6-AC、5-(dG-N(2)-基)-6-AC 和 N-(dA-8-基)-6-AC;(2)通过硝基还原和环氧化途径的组合形成反式-1,2-二羟基-1,2-二氢-6-羟基氨基苊(1,2-DHD-6-NHOH-C),生成 N-(dG-8-基)-1,2-DHD-6-AC、5-(dG-N(2)-基)-1,2-DHD-6-AC 和 N-(dA-8-基)-1,2-DHD-6-AC。如果这些 DNA 损伤在 DNA 复制发生之前没有被细胞防御机制清除,它们很可能导致突变。在这里,我们首次在 HeLa 细胞提取物的体外比较了简单的硝基还原和硝基还原与环氧化途径相结合产生的 DNA 损伤的相对核苷酸切除修复(NER)效率。我们表明,N-(dG-8-基)-1,2-DHD-6-AC 加合物比 N-(dG-8-基)-6-AC 加合物更能抵抗 NER,抗性因子约为 2 倍。此外,N-(dA-8-基)-6-AC 的修复能力要差得多,因为其 NER 效率比 N-(dG-8-基)-6-AC 加合物低约 8 倍。基于我们之前的研究和本研究,6-NC 和苯并[a]芘衍生的损伤可以按照从最易修复到最难修复的顺序排列如下:N-(dA-8-基)-6-AC>N-(dG-8-基)-1,2-DHD-6-AC>5-(dG-N(2)-基)-6-AC≈N-(dG-8-基)-6-AC≈(+)-7R,8S,9S,10S-苯并[a]芘二醇环氧化物衍生的反式-抗-苯并[a]芘-N(2)-dG 加合物。6-NC 衍生的各种损伤修复缓慢,因此它们在哺乳动物组织中的潜在持久性,部分解释了与苯并[a]芘相比,6-NC 在大鼠乳腺中的强大致癌性。