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Toxicology Research Vol - 3 Issue - 4

Toxicology Research Vol - 3 Issue - 3

Toxicology Research Vol - 3 Issue - 2

Toxicology Research Vol - 3 Issue - 1

Toxicology Research Vol -2

Toxicology Research Vol -1

Oxidative DNA damage and formalin-fixation procedures

Content Provider	Royal Society of Chemistry (RSC)
Author	Annaratone, Laura Peluso, Marco E. M. Bussolati, Gianni Giese, Roger W. Bono, Roberto Munnia, Armelle Tarocchi, Mirko
Copyright Year	2014
Abstract	Formaldehyde is the most commonly used fixative for the preparation of formalin-fixed paraffin-embedded tissues (FFPETs) in the “reduction rooms” of pathology wards. Therefore, we analysed for the generation of 3-(2-deoxy-β-D-erythro-pentafuranosyl)pyrimido[1,2-α]purin-10(3H)-one adducts (M1dG), an exocyclic DNA adduct considered to be a biomarker of oxidative stress and lipid peroxidation, in DNA extracted from the FFPETs of liver and lung samples of six C57BL/6 control mice and from the FFPETs of four cancer patients with respect to paired flash-frozen tissues by 32P-postlabeling. When the experimental animals were examined, the percentage of M1dG adducts was about 4–6 fold greater with the FFPET mouse samples as compared to flash-frozen mouse samples. Specifically, 4.75 M1dG ± 0.21 (SE) per 106 normal nucleotides (nn) were detected in the FFPET liver samples, and 1.07 M1dG ± 0.08 (SE) per 106 nn in the flash-frozen liver samples (p = 0.02). Then, 3.80 M1dG ± 0.73 (SE) per 106 nn were measured in the FFPET lung samples, and 1.02 M1dG ± 0.07 (SE) per 106 nn in the flash-frozen lung samples (p = 0.02). Also, significantly increased levels of oxidatively damaged DNA were detected in the human colon DNA from the FFPETs with respect to the flash-frozen tissues. There were 30.2 M1dG ± 7.7 (SE) per 108 nn in the colon mucosa DNA from the FFPETs and 4.4 M1dG ± 0.7 (SE) per 108 nn in the corresponding flash-frozen human tissues (p = 0.016). Formalin penetration through cell membrane components induces excessive oxidative stress, causing both direct oxidation of DNA and increased lipid peroxidation, which in turn produce M1dG adducts, a kind of DNA damage that can partially block DNA synthesis and induce error prone translesion synthesis. An excess of exocyclic DNA adducts in formalin-fixed specimens can stall DNA polymerases and contribute to the induction of artefactual sequence alterations during PRC amplification.
Starting Page	341
Ending Page	349
Page Count	9
File Format	HTM / HTML PDF
ISSN	2045452X
Volume Number	3
Issue Number	5
Journal	Toxicology Research
DOI	10.1039/c4tx00046c
Language	English
Publisher	Royal Society of Chemistry
Access Restriction	Open
Subject Keyword	PRC Oxidative stress Biomarker Cell membrane Liver Lung Colorectal cancer Formalin DNA adduct Lipid peroxidation DNA repair Formaldehyde Mucous membrane Pathology Mouse DNA Fixation (histology) Cancer
Content Type	Text
Resource Type	Article
Subject	Health, Toxicology and Mutagenesis Toxicology

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