Identification of three major DNA adducts formed by the carcinogenic air pollutant 3-nitrobenzanthrone in rat lung at the C8 and N2 position of guanine and at the N6 position of adenine
Identification of three major DNA adducts formed by the carcinogenic air pollutant 3-nitrobenzanthrone in rat lung at the C8 and N2 position of guanine and at the N6 position of adenine
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DOI：
10.1002/ijc.21622
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发表时间：
2006-05-01
影响因子：
6.4
通讯作者：
Takamura-Enya, T
中科院分区：
文献类型：
--
作者：
Arlt, VM;Schmeiser, HH;Takamura-Enya, T
3-硝基苯蒽酮（3-NBA）是一种强诱变剂和潜在的人类致癌物中确定的柴油机废气和环境空气颗粒物。以前，我们检测到的形成3-NBA衍生的DNA加合物在啮齿动物组织中的P-32-postlabeling，所有这些都是来自还原代谢产物的3-NBA结合嘌呤碱基，但这些加合物的结构鉴定尚未报道。我们现在已经制备了3-NBA衍生的DNA加合物标准品的P-32-后标记反应N-乙酰氧基-3-氨基苯并蒽酮（N-Aco-阿坝）与嘌呤核苷酸。三种脱氧鸟苷（dG）加合物已被表征为N-（2 '-脱氧鸟苷-8-基）-3-氨基苯并蒽酮-3'-磷酸（dG3 'p-C8-N-ABA），2-（2 '-脱氧鸟苷-N-2 -基）-3-氨基苯并蒽酮-3'-磷酸（dG 3 ′ p-N-2-阿坝）和2- 2 ′-（脱氧龙胆草苷-8-基）-3-氨基苯并蒽酮-3 '-磷酸（dG 3 ′ p-C8-C2-阿坝），脱氧腺苷（dA）加合物被表征为2-（2 ′-脱氧腺苷-N-6-基）-3-氨基苯并蒽酮-3 ′-磷酸（dA 3 ′ p-N-6-阿坝）。3-NBA衍生的DNA加合物在体内和体外实验形成的化学合成的加合物进行了比较。在两个独立的系统（薄层和高效液相色谱）中，大鼠肺中形成的主要3-NBA衍生的DNA加合物与dG 3 ′ p-N-2-阿坝共色谱。这也是用3-氨基苯并蒽酮（3-阿坝）（3-NBA的主要人体代谢物）处理的大鼠或小鼠组织中形成的主要加合物。同样地，dG 3 ′ p-C 8-N-ABA和dA 3 ′ p-N-6-阿坝与用3-NBA或3-阿坝处理的大鼠或小鼠的各种器官中形成的另外两种加合物共色谱，而dG 3 ′ p-C 8-C2-阿坝与体内发现的加合物中的任一种不共色谱。利用不同的体外酶系统，包括人肝微粒体和细胞质，以及纯化的和重组的酶，我们发现多种酶[NAD（P）H：醌氧化还原酶，黄嘌呤氧化酶，NADPH：细胞色素P450氧化还原酶，细胞色素P450 1A 1和1A 2，N，O-乙酰基转移酶1和2，磺基转移酶1A 1和1A 2，和髓过氧化物酶]能够在与3-NBA和/或3-阿坝孵育后催化DNA中2-（2-脱氧鸟苷-N-2-基）3-氨基苯并蒽酮，N-（2'-脱氧鸟苷-8-基）-3-氨基苯并蒽酮和2-（2'-脱氧腺苷-N-6-基）-3-氨基苯并蒽酮的形成。(c)2005 Wiley-Liss，Inc.
3-Nitrobenzanthrone (3-NBA) is a potent mutagen and potential human carcinogen identified in diesel exhaust and ambient air particulate matter. Previously, we detected the formation of 3-NBA-derived DNA adducts in rodent tissues by P-32-postlabeling, all of which are derived from reductive metabolites of 3-NBA bound to purine bases, but structural identification of these adducts has not yet been reported. We have now prepared 3-NBA-derived DNA adduct standards for P-32-postlabeling by reacting N-acetoxy-3-aminobenzanthrone (N-Aco-ABA) with purine nucleotides. Three deoxyguanosine (dG) adducts have been characterised as N-(2'-deoxy-guanosin-8-yl)-3-aminobenzanthrone-3'-phosphate (dG3'p-C8-N-ABA), 2-(2'-deoxyguanosin-N-2 -yl)-3-aminobenzanthrone-3'-phosphate (dG3'p-N-2-ABA) and 2-2'-(deoxygtianosin-8-yl)-3-aminobenzanthrone-3'-phosphate (dG3'p-C8-C2-ABA), and a deoxyadenosine (dA) adduct was characterised as 2-(2'-deoxyadenosm-N-6-yl)-3-aminobenzanthrone-3'-phosphate (dA3'p-N-6-ABA). 3-NBA-derived DNA adducts formed experimentally in vivo and in vitro were compared with the chemically synthesised adducts. The major 3-NBA-derived DNA adduct formed in rat lung cochromatographed with dG3'p-N-2-ABA in two independent systems (thin layer and high-performance liquid chromatography). This is also the major adduct formed in tissue of rats or mice treated with 3-aminobenzanthrone (3-ABA), the major human metabolite of 3-NBA. Similarly, dG3'p-C8-N-ABA and dA3'p-N-6-ABA cochromatographed with two other adducts formed in various organs of rats or mice treated either with 3-NBA or 3-ABA, whereas dG3'p-C8-C2-ABA did not cochromatograph with my of the adducts found in vivo. Utilizing different enzymatic systems in vitro, including human hepatic microsomes and cytosols, and purified and recombinant enzymes, we found that a variety of enzymes [NAD(P)H:quinone oxidoreductase, xanthine oxidase, NADPH:cytochrome P450 oxidoreductase, cytochrome P450s 1A1 and 1A2, N,O-acetyltransferases 1 and 2, sulfotransferases 1A1 and 1A2, and myeloperoxidase] are able to catalyse the formation of 2-(2-deoxyguanosin-N-2-yl)3-aminobenzanthrone, N-(2'-deoxyguanosin-8-yl)-3-aminobenzanthrone and 2-(2'-deoxyadenosin-N-6-yl)-3-aminobenzanthrone in DNA, after incubation with 3-NBA and/or 3-ABA. (c) 2005Wiley-Liss, Inc.