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伏安法测定γ辐射诱导的DNA损伤。

Voltammetric determination of gamma radiation-induced DNA damage.

作者信息

Piedade J A P, Oliveira P S C, Lopes M C, Oliveira-Brett A M

机构信息

Departamento de Química, Universidade de Coimbra, 3004-535 Coimbra, Portugal.

出版信息

Anal Biochem. 2006 Aug 1;355(1):39-49. doi: 10.1016/j.ab.2006.05.022. Epub 2006 Jun 5.

Abstract

Homopolydeoxyribonucleotides, poly[dGuo], poly[dAdo], poly[dThd], and poly[dCyd], and calf thymus single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA) aqueous solutions previously exposed to gamma radiation doses between 2 and 35 Gy, were studied by differential pulse voltammetry using a glassy carbon electrode. The interpretation of the voltammetric data was also supported by the electrophoretic migration profile obtained for the same ssDNA and dsDNA gamma-irradiated samples by nondenaturing agarose gel electrophoresis. The generation of 8-oxo-7,8-dihydroguanine, 2,8-dihydroxyadenine, 5-formyluracil, base-free sites, and single- and double-stranded breaks in the gamma-irradiated DNA samples was detected voltammetrically, with the amount depending on the irradiation time. It was found that the current peaks obtained for 8-oxoguanine increase linearly with the radiation dose applied to the nucleic acid sample, and values between 8 and 446 8-oxo-7,8-dihydroguanine (8-oxoGua) per 10(6) guanines per Gy were obtained according to the nucleic acid sample. The results showed that voltammetry can be used for monitoring and simultaneously characterizing different kinds of DNA damage caused by gamma radiation exposure.

摘要

使用玻碳电极通过差分脉冲伏安法研究了均聚脱氧核糖核苷酸、聚[dGuo]、聚[dAdo]、聚[dThd]和聚[dCyd],以及先前暴露于2至35 Gyγ辐射剂量下的小牛胸腺单链DNA(ssDNA)和双链DNA(dsDNA)水溶液。通过非变性琼脂糖凝胶电泳对相同的ssDNA和dsDNAγ辐照样品获得的电泳迁移图谱也支持了伏安数据的解释。通过伏安法检测了γ辐照DNA样品中8-氧代-7,8-二氢鸟嘌呤、2,8-二羟基腺嘌呤、5-甲酰基尿嘧啶、无碱基位点以及单链和双链断裂的产生,其数量取决于辐照时间。发现8-氧代鸟嘌呤的电流峰随施加于核酸样品的辐射剂量线性增加,根据核酸样品,每Gy每10(6)个鸟嘌呤中8-氧代-7,8-二氢鸟嘌呤(8-oxoGua)的值在8至446之间。结果表明,伏安法可用于监测并同时表征由γ辐射暴露引起的不同类型的DNA损伤。

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