Identification and quantification of DNA adducts in the oral tissues of mice treated with the environmental carcinogen dibenzo[a,l]pyrene by HPLC-MS/MS.
Identification and quantification of DNA adducts in the oral tissues of mice treated with the environmental carcinogen dibenzo[a,l]pyrene by HPLC-MS/MS.
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DOI：
10.1021/tx200188j
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发表时间：
2011-08-15
影响因子：
4.1
通讯作者：
El-Bayoumy K
中科院分区：
文献类型：
--
作者：
Zhang SM;Chen KM;Aliaga C;Sun YW;Lin JM;Sharma AK;Amin S;El-Bayoumy K
吸烟是口腔癌的主要原因之一。二苯并[a,l]芘 (DB[a,l]P) 是一种环境污染物和烟草烟雾成分，是迄今为止在多种动物模型（靶器官：皮肤、肺、卵巢和乳腺组织）中测试的致癌性最强的多环芳烃 (PAH)。我们最近证明 DB[a,l]P 也能够诱导小鼠口腔癌；然而，其在小鼠口腔中对最终遗传毒性代谢物二苯并[a,l]芘-11,12-二氢二醇-13,14-环氧化物（DB[a,l]PDE）的代谢激活尚未得到研究。在这里，我们开发了一种液相色谱-串联质谱 (LC-MS/MS) 方法来检测和定量接受 DB[a,l]P 治疗的小鼠口腔组织中的 (±)-抗 DB[a,l]PDE-dA 加合物。合成[15N5]-(±)-抗DB[a,l]PDE-N6-dA加合物作为内标。通过 MS、NMR 和 CD 分析对立体异构加合物进行了表征。以 100 µg 消化 DNA 为基质，该方法的检测限为 8 fmol。将DB[a,l]P（每天240 nmol，持续2天）直接施用到口腔后，在小鼠口腔组织中检测到两种加合物并鉴定为(−)-抗顺式和(−)-抗反式-DB[a,l]PDE-dA，表明DB[a,l]P在该靶器官中主要代谢为(−)-抗-DB[a,l]PDE。我们还比较了将 DB[a,l]P（24 nmol，每周 3 次，持续 5 周）直接施用到小鼠口腔后，加合物的形成和去除随时间的变化。在最后一次给药后 48 小时、1、2 和 4 周对加合物进行定量。加合物的最高水平出现在 48 小时，随后逐渐下降。每个时间点(-)-抗反式加合物的水平(fmol/μg DNA)(4.03±0.27至1.77±0.25)显着高于(-)-抗顺-DB[a,l]PDE-dA加合物(1.63±0.42至0.72±0.04)(p < 0.005)。本文的结果表明，(−)-抗 DB[a,l]PDE-dA 加合物的形成和持续存在可能部分促进 DB[a,l]P 诱导的口腔癌发生。
Tobacco smoking is one of the leading causes for oral cancer. Dibenzo[a,l]pyrene (DB[a,l]P), an environmental pollutant and a tobacco smoke constituent, is the most carcinogenic polycyclic aromatic hydrocarbon (PAH) tested to date in several animal models (target organs: skin, lung, ovary and mammary tissues). We have recently demonstrated that DB[a,l]P is also capable of inducing oral cancer in mice; however its metabolic activation to the ultimate genotoxic metabolite dibenzo[a,l]pyrene-11,12-dihydrodiol-13,14-epoxide (DB[a,l]PDE) in mouse oral cavity has not been examined. Here we developed a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to detect and quantify (±)-anti-DB[a,l]PDE-dA adducts in oral tissues of mice treated with DB[a,l]P. [15N5]-(±)-anti-DB[a,l]PDE-N6-dA adducts were synthesized as internal standards. The stereoisomeric adducts were characterized by MS, NMR and CD analysis. The detection limit of the method is 8 fmol with 100 µg digested DNA as the matrix. Two adducts were detected and identified as (−)-anti-cis and (−)-anti-trans-DB[a,l]PDE-dA in the oral tissues of mice following direct application of DB[a,l]P (240 nmol per day, for 2 days) into the oral cavity, indicating that DB[a,l]P is predominantly metabolized into (−)-anti-DB[a,l]PDE in this target organ. We also compared the formation and removal of adducts as a function of time, following direct application of DB[a,l]P (24 nmol, 3 times per week for 5 weeks) into the oral cavity of mice. Adducts were quantified at 48 h, 1, 2 and 4 weeks after the last dose. Maximal levels of adducts occurred at 48 h, followed by a gradual decrease. The levels (fmol/µg DNA) of (−)-anti-trans adducts (4.03±0.27 to 1.77±0.25) are significantly higher than (−)-anti-cis-DB[a,l]PDE-dA adduct (1.63±0.42 to 0.72±0.04) at each time point (p < 0.005). The results presented here indicate that the formation and persistence of (−)-anti-DB[a,l]PDE-dA adducts may, in part, contribute to the initiation of DB[a,l]P-induced oral carcinogenesis.
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影响因子：
4.1
作者：
Cai, Yuqin;Ding, Shuang;Broyde, Suse
通讯作者：
Broyde, Suse
影响因子：
4.1
作者：
Ruan, Q;Kolbanovskiy, A;Geacintov, NE
通讯作者：
Geacintov, NE
影响因子：
2.9
作者：
Cai, Yuqin;Wang, Lihua;Broyde, Suse
通讯作者：
Broyde, Suse
影响因子：
4.7
作者：
Prahalad, AK;Ross, JA;Mass, MJ
通讯作者：
Mass, MJ
影响因子：
15
作者：
AGARWAL, SK;SAYER, JM;JERINA, DM
通讯作者：
JERINA, DM