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Updated: Aug 5, 2026
Measuring DNA Damage and Repair in Mouse Splenocytes After Chronic In Vivo Exposure to Very Low Doses of Beta- and Gamma-Radiation
Published on: July 3, 2015
DNA damage measured by liquid chromatography-mass spectrometry in mouse fibroblast cells exposed to oxidative stress
Jean B Dawidzik1, Helen B Patrzyc, Herbert Iijima
1Department of Molecular and Cellular Biophysics, Roswell Park Cancer Institute, Elm and Carlton Streets, Buffalo, NY 14263, USA.
Abstract:
Oxidative DNA damage can result from environmental factors, such as radiation, as well as from the untoward consequences of normal metabolic processes. It is of interest to assay oxidative DNA damage in cells and tissues because this damage has been implicated in human disease, particularly cancer. Eleven indicators of oxidative DNA damage have been measured by Liquid Chromatography-Mass Spectrometry (LC-MS) in DNA extracted from cells exposed to oxidative stress. Mouse fibroblast cells were exposed to hydrogen peroxide and to UVC light and to the combined action of both agents. Significant increases of the 8-oxo-7,8-dihydropurine lesions over background were detected. Significant increases of the formamido lesions resulting from breakdown of pyrimidine bases were also observed. Of special interest was the observation of double lesions, tandem combinations of both aforementioned lesions, in cells exposed to oxidative stress.
Insights
This study measured oxidative DNA damage using Liquid Chromatography-Mass Spectrometry (LC-MS). Researchers found increased DNA lesions, including novel double lesions, in cells exposed to oxidative stress.
Area of Science:
- Biochemistry
- Molecular Biology
- Genetics
Background:
- Oxidative DNA damage arises from environmental factors (e.g., radiation) and normal metabolism.
- This damage is linked to human diseases, notably cancer.
- Assaying oxidative DNA damage is crucial for understanding disease mechanisms.
Purpose of the Study:
- To quantify oxidative DNA damage in cells under stress.
- To identify specific DNA lesions resulting from oxidative insults.
- To investigate the occurrence of combined DNA lesions.
Main Methods:
- Mouse fibroblast cells were subjected to oxidative stress using hydrogen peroxide and UVC light.
- Eleven indicators of oxidative DNA damage were measured.
- Liquid Chromatography-Mass Spectrometry (LC-MS) was employed for precise quantification.
Main Results:
- Significant increases in 8-oxo-7,8-dihydropurine lesions were detected.
- Elevated levels of formamido lesions, from pyrimidine base breakdown, were observed.
- Novel double lesions, combining both types of damage, were identified in stressed cells.
Conclusions:
- Oxidative stress induces distinct types of DNA damage, including purine and pyrimidine lesions.
- The formation of double lesions represents a significant finding in oxidative DNA damage research.
- These findings contribute to understanding the molecular basis of diseases linked to DNA damage.
