2001
DOI: 10.1667/0033-7587(2001)155[0832:dooiic]2.0.co;2
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Determination of 8-Oxoguanine in Individual Cell Nucleus of Gamma-Irradiated Mammalian Cells

Abstract: 8-Oxoguanine, through its ability to mispair bases other than cytosine, is assumed to be one of the most potent premutagenic lesions in nuclear DNA damaged by reactive oxygen radicals. In this study, we examine whether the presence of residual 8-oxoguanine can be detected in mammalian cells after exposure to ionizing radiation. MOLT-4 human leukemia cells and CHO-K1 Chinese hamster cells were acutely irradiated in vitro with 0, 0.2, 0.4, 0.6 and 1.0 Gy gamma radiation at room temperature. The amounts of 8-oxog… Show more

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Cited by 22 publications
(21 citation statements)
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“…ROS production and/or complex chemical alterations of neighbouring nucleotides are indeed frequently associated with conventional clastogenic treatments and complicate result interpretation due to unrelated phenomenon such as kinase activations and base excision repair (BER) engagement. 44 Second, using ChIP followed by classical Q-PCR analysis allows investigation of DSB repair protein recruitment at a molecular level, which is sometimes critical in order to observe local events undetectable by conventional immunofluorescence. For example, we could see a clear recruitment of XRCC4 by ChIP, while no IRIF association was detectable by IF.…”
Section: Characterizing the Chromatin Structure And Function In Dsb Rmentioning
confidence: 99%
“…ROS production and/or complex chemical alterations of neighbouring nucleotides are indeed frequently associated with conventional clastogenic treatments and complicate result interpretation due to unrelated phenomenon such as kinase activations and base excision repair (BER) engagement. 44 Second, using ChIP followed by classical Q-PCR analysis allows investigation of DSB repair protein recruitment at a molecular level, which is sometimes critical in order to observe local events undetectable by conventional immunofluorescence. For example, we could see a clear recruitment of XRCC4 by ChIP, while no IRIF association was detectable by IF.…”
Section: Characterizing the Chromatin Structure And Function In Dsb Rmentioning
confidence: 99%
“…It had previously been shown by high-performance liquid chromatography (HPLC) that H 2 O 2 induces 8-oxo-dG, a DNA damage marker [16]. Exposure of quiescent WI-26VA4 cells, another SV-40 transformed epithelial cell line, to H 2 O 2 had no effect on the number of 8-oxo-dG-positive cells (Fig.…”
Section: Resultsmentioning
confidence: 82%
“…We used an FITC-conjugated substrate to detect 8-oxo-dG and analyzed this by LSC at the single cell level. Traditional methods using HPLC [16] can assess the total amount of DNA damage per genomes whereas LSC indicates DNA damage at each individual cell. The detection of 8-oxo-dG by fluorescence in a flow cytometer [30] or by LSC might be useful tools for estimating DNA damage at the single cell level.…”
Section: Discussionmentioning
confidence: 99%
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“…Next, we examined the level of 8-oxoG DNA lesions in the mutant cells by using FITC-avidin/PI fluorescence microscopy. As FITC-avidin specifically labels DNA lesions while PI labels all DNA, the intensity ratio of stained FITC to PI reflects the level of 8-oxoG DNA lesions in the cells (41). This technique was successfully used in our lab to examine other oxidative-stress-related mutants of H. pylori (42,43).…”
Section: Resultsmentioning
confidence: 99%