Adenine-containing DNA-DNA cross-links of antitumor nitrogen mustards
Adenine-containing DNA-DNA cross-links of antitumor nitrogen mustards
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DOI：
10.1021/tx0499463
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发表时间：
2004-07-01
影响因子：
4.1
通讯作者：
Tretyakova, N
中科院分区：
文献类型：
--
作者：
Balcome, S;Park, S;Tretyakova, N
氮芥（NM）是治疗淋巴瘤、白血病、多发性骨髓瘤和卵巢癌的有用化疗剂。 NM 的抗肿瘤活性归因于它们交联 DNA 双链的能力。由此产生的双功能损伤如果不修复，会抑制 DNA 复制和转录，最终导致细胞周期停滞、细胞凋亡和肿瘤生长的抑制。据报道，NM 的主要双功能 DNA 损伤涉及 5'-GNC 序列相对链中的远端鸟嘌呤碱基。在本工作中，证明了双链 DNA 中 N,N-双（2-氯乙基）甲胺（氮芥）的鸟嘌呤-腺嘌呤和腺嘌呤-腺嘌呤加合物的形成。二氯乙胺的鸟嘌呤-腺嘌呤交联被鉴定为N-(2-[N3-腺苷基]乙基)-N-(2-[N7-鸟苷基]乙基)甲胺(N3A-N7G-EMA)、N-(2-[N1-腺苷基]乙基)-N-(2-[N7-鸟苷基]乙基)甲胺，和N-(2-[N-6-腺基]乙基)-N-(2-[N7-脒基]乙基)甲胺。所有三种加合物均产生链间，而 N3A-N7G-EMA 是主要的链内 G-A 交联。常见的腺嘌呤-腺嘌呤二氯乙胺损伤具有N,N-双(2-[N3-腺苷基]乙基)甲胺(bis-N3A-EMA)的结构。 DNA 来源的损伤与独立制备的真实标准品具有相同的 HPLC 保留时间、UV 光谱和 MS/MS 碎片模式。在用增加量的氮芥处理的小牛胸腺DNA中以浓度依赖性方式产生bis-N3A-EMA损伤。此外，使用 HPLC-ESI-MS/MS 分析证明，用芳香氮芥、N,N-双(2-氯乙基)-对氨基苯基丁酸和 L-苯基丙氨酸芥处理的 DNA 中类似的 N3-N3 腺嘌呤损伤的形成。交联腺嘌呤-腺嘌呤损伤的存在可以解释 NM 在缺乏 N3-烷基腺嘌呤糖基化酶的细胞中增强的细胞毒性和致突变性。
Nitrogen mustards (NMs) are useful chemotherapeutic agents in the treatment of lymphoma, leukemia, multiple myeloma, and ovarian carcinoma. The antitumor activity of NMs has been attributed to their ability to cross-link the twin strands of DNA. The resulting bifunctional lesions, if not repaired, can inhibit DNA replication and transcription, eventually leading to cell cycle arrest, apoptosis, and the inhibition of tumor growth. The predominant bifunctional DNA lesions of NM have been reported to involve the distal guanine bases in the opposite strands of 5'-GNC sequences. In the present work, the formation of guanine-adenine and adenine-adenine adducts of N,N-bis(2-chloroethyl)methylamine (mechlorethamine) in double-stranded DNA is demonstrated. Guanine-adenine cross-links of mechlorethamine were identified as N-(2-[N3-adenyl]ethyl)-N-(2-[N7-guanyl]ethyl)methylamine (N3A-N7G-EMA), N-(2-[N1-adenyl]ethyl)-N-(2-[N7-guanyl]ethyl)methylamine, and N-(2-[N-6-adenyl]ethyl)-N-(2-[N7-guanyl]ethyl)methylamine. All three adducts were produced interstrand, while N3A-N7G-EMA was the dominant intrastrand G-A cross-link. The prevalent adenine-adenine mechlorethamine lesions have the structure of N,N-bis(2-[N3-adenyl]ethyl)methylamine (bis-N3A-EMA). DNA-derived lesions have the same HPLC retention times, UV spectra, and MS/MS fragmentation patterns as the authentic standards prepared independently. bis-N3A-EMA lesions were produced in a concentration-de pendent manner in calf thymus DNA treated with increasing amounts of mechlorethamine. Furthermore, HPLC-ESI-MS/MS analysis was used to demonstrate the formation of analogous N3-N3 adenine lesions in DNA treated with aromatic nitrogen mustards, N,N-bis(2-chloroethyl)-p-aminophenylbutyric acid and L-phenyl-alanine mustard. The presence of cross-linked adenine-adenine lesions may explain the enhanced cytotoxicity and mutagenicity of NMs in cells deficient in N3-alkyladenine glycosylase.