An approach based on ultra-high pressure liquid chromatography–tandem mass spectrometry to quantify O6-methyl and O6-carboxymethylguanine DNA adducts in intestinal cell lines

Bussche, Julie Vanden; Moore, Sharon A.; Pasmans, Frank; Kuhnle, Gunter; Vanhaecke, Lynn

doi:10.1016/j.chroma.2012.07.040

Cited by 29 publications

(66 citation statements)

References 23 publications

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“…The NOC-specific DNA adduct O 6 -C-MeG was quantified using U-HPLC-MS/MS [26]. In brief, 182 µl of filter sterilized sample was incubated for 18 hours at 37°C with 100 µg calf thymus (CT-)DNA.…”

Section: Methodsmentioning

confidence: 99%

Fat Content and Nitrite-Curing Influence the Formation of Oxidation Products and NOC-Specific DNA Adducts during In Vitro Digestion of Meat

et al. 2014

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The effects of fat content and nitrite-curing of pork were investigated on the formation of cytotoxic and genotoxic lipid oxidation products (malondialdehyde, 4-hydroxy-2-nonenal, volatile simple aldehydes), protein oxidation products (protein carbonyl compounds) and NOC-specific DNA adducts (O6-carboxy-methylguanine) during in vitro digestion. The formation of these products during digestion is suggested to be responsible for the association between red meat and processed meat consumption and colorectal cancer risk. Digestion of uncured pork to which fat was added (total fat content 5 or 20%), resulted in significantly higher lipid and protein oxidation in the mimicked duodenal and colonic fluids, compared to digestion of pork without added fat (1% fat). A higher fat content also significantly favored the formation of O6-carboxy-methylguanine in the colon. Nitrite-curing of meat resulted in significantly lower lipid and protein oxidation before and after digestion, while an inconsistent effect on the formation of O6-carboxy-methylguanine was observed. The presented results demonstrate that haem-Fe is not solely responsible for oxidation and nitrosation reactions throughout an in vitro digestion approach but its effect is promoted by a higher fat content in meat.

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Section: Methodsmentioning

confidence: 99%

Fat Content and Nitrite-Curing Influence the Formation of Oxidation Products and NOC-Specific DNA Adducts during In Vitro Digestion of Meat

et al. 2014

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“…14 Preferential formation of O 6 -CMdG over O 6 -MedG was observed for isolated DNA treated with various DNA carboxymethylating agents and for human cells treated with diazoacetate. 10,17 These findings, along with the fact that O 6 -MedG induces predominantly GC→AT transition mutation, 18 indicate that DNA carboxymethylation, rather than methylation, may contribute to the mutations of p53 gene found in gastrointestinal cancer. 14 Recent replication and transcription studies with the use of plasmids housing a site-specifically inserted DNA lesion showed that O 4 - and N 3-carboxymethylthymidine could substantially stall DNA replication and transcription in cells, and induce mutations in these processes.…”

Section: Introductionmentioning

confidence: 99%

“…24,25 In this regard, solid-phase extraction followed by LC-MS/MS analysis on a triple-quadrupole mass spectrometer was previously employed for the quantification of O 6 -CMdG and O 6 -MedG, directly or after converting them to the corresponding modified nucleobases with acid hydrolysis at elevated temperature. 17,26 Although neither approach provided adequate sensitivity for detecting these modified nucleosides in human urine samples, 26 LC-MS/MS analysis of the acid-hydrolyzed samples facilitated the successful detection of O 6 -CMdG and O 6 -MedG in DNA isolated from cultured human cells treated with diazoacetate. 17 …”

Section: Introductionmentioning

confidence: 99%

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Quantification of Azaserine-Induced Carboxymethylated and Methylated DNA Lesions in Cells by Nanoflow Liquid Chromatography-Nanoelectrospray Ionization Tandem Mass Spectrometry Coupled with the Stable Isotope-Dilution Method

Wang

et al. 2016

Anal. Chem.

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Humans are exposed to N-nitroso compounds through environmental exposure and endogenous metabolism. Some N-nitroso compounds can be metabolically activated to yield diazoacetate, which is known to induce DNA carboxymethylation. DNA lesion measurement remains one of the core tasks in toxicology and in evaluating human health risks associated with carcinogen exposure. In this study, we developed a highly sensitive nano-flow liquid chromatography-nanoelectrospray ionization-multi-stage tandem mass spectrometry (nLC-nESI-MS3) method for the simultaneous quantification of O6-carboxymethyl-2′-deoxyguanosine (O6-CMdG), O6-methyl-2′-deoxyguanosine (O6-MedG), and N6-carboxymethyl-2′-deoxyadenosine (N6-CMdA). We were able to measure the levels of these three lesions with the use of low-microgram quantities of DNA from cultured human skin fibroblasts and human colorectal carcinoma cells treated with azaserine, a DNA carboxymethylating agent. Our results revealed that the levels of O6-CMdG and O6-MedG increased when the dose of azaserine was increased from 0 to 450 µM. We, however, did not observe an apparent dose-dependent induction of N6-CMdA, suggesting the presence of repair mechanism(s) for the rapid clearance of this lesion in cells. This is the first report about the application of nLC-nESI-MS3 technique for the simultaneous quantification of O6-CMdG, O6-MedG and N6-CMdA. The method reported here will be useful for the future investigations about the repair of the carboxymethylated DNA lesions and about the implications of these lesions in carcinogenesis.

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“…The demonstrated dose response effect was to be expected for KDA, CMG, since the methylation and carboxymethylation of deoxyguanosine at the O 6 position by nitrosated glycine derivatives have been documented in the past[19,36]. The demonstrated formation of M1G in CT-DNA by attack of the G base by the highly reactive lipid peroxidation product MDA has also been reported extensively in literature, with M1G…”

mentioning

confidence: 68%

High resolution mass spectrometry based profiling of diet-related deoxyribonucleic acid adducts

Hemeryck

Decloedt

Bussche

et al. 2015

Analytica Chimica Acta

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Exposure of DNA to endo- and exogenous DNA binding chemicals can result in the formation of DNA adducts and is believed to be the first step in chemically induced carcinogenesis. DNA adductomics is a relatively new field of research which studies the formation of known and unknown DNA adducts in DNA due to exposure to genotoxic chemicals. In this study, a new UHPLC-HRMS(/MS)-based DNA adduct detection method was developed and validated. Four targeted DNA adducts, which all have been linked to dietary genotoxicity, were included in the described method; O(6)-methylguanine (O(6)-MeG), O(6)-carboxymethylguanine (O(6)-CMG), pyrimidopurinone (M1G) and methylhydroxypropanoguanine (CroG). As a supplementary tool for DNA adductomics, a DNA adduct database, which currently contains 123 different diet-related DNA adducts, was constructed. By means of the newly developed method and database, all 4 targeted DNA adducts and 32 untargeted DNA adducts could be detected in different DNA samples. The obtained results clearly demonstrate the merit of the described method for both targeted and untargeted DNA adduct detection in vitro and in vivo, whilst the diet-related DNA adduct database can distinctly facilitate data interpretation.

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An approach based on ultra-high pressure liquid chromatography–tandem mass spectrometry to quantify O6-methyl and O6-carboxymethylguanine DNA adducts in intestinal cell lines

Cited by 29 publications

References 23 publications

Fat Content and Nitrite-Curing Influence the Formation of Oxidation Products and NOC-Specific DNA Adducts during In Vitro Digestion of Meat

Fat Content and Nitrite-Curing Influence the Formation of Oxidation Products and NOC-Specific DNA Adducts during In Vitro Digestion of Meat

Quantification of Azaserine-Induced Carboxymethylated and Methylated DNA Lesions in Cells by Nanoflow Liquid Chromatography-Nanoelectrospray Ionization Tandem Mass Spectrometry Coupled with the Stable Isotope-Dilution Method

High resolution mass spectrometry based profiling of diet-related deoxyribonucleic acid adducts

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