Determination of malondialdehyde-induced DNA damage in human tissues using an immunoslot blot assay

被引:96
作者
Leuratti, C
Singh, R
Lagneau, C
Farmer, PB
Plastaras, JP
Marnett, LJ
Shuker, DEG
机构
[1] Univ Leicester, MRC Toxicol Unit, Leicester LE1 9HN, Leics, England
[2] Vanderbilt Univ, Dept Biochem, Nashville, TN 37232 USA
关键词
D O I
10.1093/carcin/19.11.1919
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
Malondialdehyde (MDA) is a product of lipid peroxidation and prostaglandin biosynthesis. It is mutagenic and carcinogenic and the major adduct formed by reaction with DNA, a highly fluorescent pyrimidopurinone (M-1-dG), has been detected in healthy human liver and leukocyte DNA, Analytical methods used so far for the detection of M1-dG have not been applied to a large number of individuals or variety of samples. Often, only a few mu g of DNA from human tissues are available for analysis and a very sensitive assay is needed in order to detect background levels of M1-dG in very small amounts of DNA, In this paper, the development of an immunoslot blot (ISB) assay for the measurement of M-1-dG in 1 mu g of DNA is described. The limit of detection of the assay is 2.5 adducts per 10(8) bases, A series of human samples were analysed and levels of 5.6-9.5 (rt = 8) and 3.1-64.3 (n = 42) of M1-dG per 108 normal bases were detected in white blood cell and gastric biopsy DNA, respectively. Results on four human samples were compared with those obtained using an HPLC/P-32-post-labelling (HPLC/PPL) method previously developed and indicated a high correlation between M1-dG levels measured by the two assays. The advantages of ISB over other assays including HPLC/PPL, such as the possibility of analysing 1 mu g DNA/sample and the fact that it is less time-consuming and laborious, means that it can be more easily used for routine analysis of a large number of samples in biomonitoring studies.
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页码:1919 / 1924
页数:6
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