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一种评估DNA损伤的新方法:胸腺嘧啶乙二醇损伤的测量。

A novel approach to DNA damage assessments: measurement of the thymine glycol lesion.

作者信息

Bailey Douglas T, DeFedericis Han-Chun C, Greene Kellee F, Iijima Herbert, Budzinski Edwin E, Patrzyc Helen B, Dawidzik Jean B, Box Harold C

机构信息

Roswell Park Cancer Institute, Buffalo, New York 14263, USA.

出版信息

Radiat Res. 2006 Apr;165(4):438-44. doi: 10.1667/rr3534.1.

Abstract

A different approach to the measurement of DNA damage has been developed based on the fact that many lesions can be excised from DNA in the form of modified dinucleoside monophosphates. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is used in conjunction with isotopically labeled internal standards to quantify the lesion. The method has several advantages, including high sensitivity for the detection of dinucleoside monophosphates. The method was applied to the measurement of the 5,6-dihydroxy-5,6-dihydrothymine (thymine glycol) lesion in the DNA of mouse fibroblast cells exposed in culture to various treatments including ionizing radiation, UVC light and buthionine sulfoximine (BSO), an inhibitor of glutathione synthesis. The application of the method to the measurement of other DNA lesions is discussed.

摘要

基于许多损伤能够以修饰的二磷酸核苷单磷酸的形式从DNA中切除这一事实,已经开发出一种不同的DNA损伤测量方法。液相色谱-串联质谱法(LC-MS/MS)与同位素标记的内标物一起用于定量损伤。该方法具有几个优点,包括对二磷酸核苷单磷酸检测的高灵敏度。该方法被应用于测量在培养中暴露于各种处理(包括电离辐射、UVC光和谷胱甘肽合成抑制剂丁硫氨酸亚砜胺(BSO))的小鼠成纤维细胞DNA中的5,6-二羟基-5,6-二氢胸腺嘧啶(胸腺嘧啶二醇)损伤。还讨论了该方法在测量其他DNA损伤中的应用。

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