Hydantoin derivative formation from oxidation of 7,8-dihydro-8-oxo-2′-deoxyguanosine (8-oxodG) and incorporation of 14C-labeled 8-oxodG into the DNA of human breast cancer cells

Hah, Sang Soo; Kim, Hyung M.; Sumbad, Rhoda A.; Henderson, Paul

doi:10.1016/j.bmcl.2005.05.113

Cited by 36 publications

(32 citation statements)

References 31 publications

(4 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…T24 cells were dosed with 0.2 µM of carboplatin containing 300 dpm (2.16 pmol) of [ 14 C]carboplatin in 5 mL of the growth media. After a 2-day incubation, the radiocarbon contents of whole cell lysis mixtures and extracted DNA were measured by AMS according to published procedures (17). All T24 dosing experiments were performed in triplicate.…”

Section: Methodsmentioning

confidence: 99%

“…AMS is a sensitive and precise technology for tracing 14 Clabeled compounds in biological systems that affords subpicomole to zeptomole 14 C per milligram of total carbon sensitivity with a few percent precision (15)(16)(17). Because only carboplatin bound to DNA in the form of monoadducts contains radiocarbon, as shown in Scheme 1, AMS can only detect carboplatin monoadducts from DNA samples.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Kinetics of Carboplatin−DNA Binding in Genomic DNA and Bladder Cancer Cells As Determined by Accelerator Mass Spectrometry

Hah

Stivers

White

et al. 2006

Chem. Res. Toxicol.

Self Cite

101

View full text Add to dashboard Cite

Cisplatin and carboplatin are platinum-based drugs that are widely used in cancer chemotherapy. The cytotoxicity of these drugs is mediated by platinum-DNA monoadducts and intra-and interstrand diadducts, which are formed following uptake of the drug into the nucleus of cells. The pharmacodynamics of carboplatin display fewer side effects than for cisplatin, albeit with less potency, which may be due to differences in rates of DNA adduct formation. We report the use of accelerator mass spectrometry (AMS), a sensitive detection method often used for radiocarbon quantitation, to measure both the kinetics of [ 14 C]carboplatin-DNA adduct formation with genomic DNA and drug uptake and DNA binding in T24 human bladder cancer cells. Only carboplatin-DNA monoadducts contain radiocarbon in the platinated DNA, which allowed for calculation of kinetic rates and concentrations within the system. The percent of radiocarbon bound to salmon sperm DNA in the form of monoadducts was measured by AMS over 24 h. Knowledge of both the starting concentration of the parent carboplatin and the concentration of radiocarbon in the DNA at a variety of time points allowed calculation of the rates of Pt-DNA monoadduct formation and conversion to toxic cross-links. Importantly, the rate of carboplatin-DNA monoadduct formation was approximately 100-fold slower than that reported for the more potent cisplatin analogue, which may explain the lower toxicity of carboplatin. T24 human bladder cancer cells were incubated with a subpharmacological dose of [ 14 C]carboplatin, and the rate of accumulation of radiocarbon in the cells and nuclear DNA was measured by AMS. The lowest concentration of radiocarbon measured was approximately 1 amol/10 µg of DNA. This sensitivity may allow the method to be used for clinical applications.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Kinetics of Carboplatin−DNA Binding in Genomic DNA and Bladder Cancer Cells As Determined by Accelerator Mass Spectrometry

Hah

Stivers

White

et al. 2006

Chem. Res. Toxicol.

Self Cite

101

View full text Add to dashboard Cite

show abstract

“…Recently, 8-oxodG has been characterized as a chemically unstable intermediate because of a low oxidation potential compared with the parent dG (9,10). The oxidation of 8-oxodG produces several 2Ј-deoxynucleosides, including derivatives of cyanuric acid, oxaluric acid, oxazalone, and at least three hydantoin derivatives [for structures see supporting information (SI) Fig.…”

mentioning

confidence: 99%

“…We recently reported the synthesis of [ 14 C]8-oxodG and preliminary measurement of its incorporation into the DNA of MCF-7 cells by using AMS (9). Thus, 8-oxodG metabolism is a useful model to test the utility of AMS for metabolic flux analysis.…”

mentioning

confidence: 99%

Measurement of 7,8-dihydro-8-oxo-2′-deoxyguanosine metabolism in MCF-7 cells at low concentrations using accelerator mass spectrometry

Hah

Mundt

Kim

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

View full text Add to dashboard Cite

Growing evidence suggests that oxidative damage to cells generates mutagenic 7,8-dihydro-8-oxo-2 -deoxyguanosine (8-oxodG), which may initiate diseases related to aging and carcinogenesis. Kinetic measurement of 8-oxodG metabolism and repair in cells has been hampered by poor assay sensitivity and by difficulty characterizing the flux of oxidized nucleotides through the relevant metabolic pathways. We report here the development of a sensitive and quantitative approach to characterizing the kinetics and metabolic sources of 8-oxodG in MCF-7 human breast cancer cells by accelerator mass spectrometry. We observed that [ 14 C]8-oxodG at medium concentrations of up to 2 pmol/ml was taken up by MCF-7 cells, phosphorylated to mono-, di-, and triphosphate derivatives, and incorporated into DNA. Oxidative stress caused by exposure of the cells to 17␤-estradiol resulted in a reduction in the rate of [ 14 C]8-oxodG incorporation into DNA and an increase in the ratio of 8-oxodG monophosphate (8-oxodGMP) to 8-oxodG triphosphate (8-oxodGTP) in the nucleotide pool. 17␤-Estradiolinduced oxidative stress up-regulated the nucleotide pool cleansing enzyme MTH1 and possibly other Nudix-related pyrophosphohydrolases. These data support the conclusion that 8-oxodGTP is formed in the nucleotide pool by both 8-oxodG metabolism and endogenous reactive oxygen species. The metabolism of 8-oxodG to 8-oxodGTP, followed by incorporation into DNA is a mechanism by which the cellular presence of this oxidized nucleoside can lead to mutations.DNA repair ͉ nucleoside metabolism ͉ oxidative stress ͉ breast cancer

show abstract

“…Recently, it was reported that 8-OH-dG in cell culture medium is incorporated into the DNA of human cultured cells possibly by a salvage pathway (4). The induction of apoptosis due to the incorporation of 8-OH-dG into DNA and its accumulation in OGG1-deˆcient human cells has also been reported (5).…”

Section: Introductionmentioning

confidence: 99%

Detection of Genotoxic Nucleosides, 8-Hydroxydeoxyguanosine, 8-Hydroxyguanosine and Free Base 8-Hydroxyguanine, in Fish Food Products

Kawai

Svoboda

Kasai

2006

Genes and Environment

View full text Add to dashboard Cite

8-Hydroxydeoxyguanosine (8-OH-dG) is a DNA base modiˆcation induced by reactive oxygen species (ROS), and is analyzed in cellular DNA and urine as a marker of oxidative stress. We now report that 8-OH-dG, the ribonucleoside 8-OH-Guo and the free base 8-OH-Gua were detected inˆsh food products, such as salted dried sardines (Maruboshi) and dried smallˆshes (Iriko and Shirasu), which are often consumed in Japan. Water extracts of theseˆsh products were analyzed by an HPLC-ECD method, using anion exchange-and reverse phasecolumns. Various amounts of 8-OH-Gua (25 ng-22 mg/g), 8-OH-Guo (28-406 ng/g) and 8-OH-dG (5-60 ng/g) were detected in these foods. The amounts of 8-OH-dG and 8-OH-Guo increased upon heating or broiling of theseˆsh products. This is theˆrst report on the presence of genotoxic nucleoside analogues in food. These results provide the warnings that these nucleosides analogues might be food carcinogens, because they are incorporated into DNA by a salvage pathway and show genotoxicity.

show abstract

Hydantoin derivative formation from oxidation of 7,8-dihydro-8-oxo-2′-deoxyguanosine (8-oxodG) and incorporation of 14C-labeled 8-oxodG into the DNA of human breast cancer cells

Cited by 36 publications

References 31 publications

Kinetics of Carboplatin−DNA Binding in Genomic DNA and Bladder Cancer Cells As Determined by Accelerator Mass Spectrometry

Kinetics of Carboplatin−DNA Binding in Genomic DNA and Bladder Cancer Cells As Determined by Accelerator Mass Spectrometry

Measurement of 7,8-dihydro-8-oxo-2′-deoxyguanosine metabolism in MCF-7 cells at low concentrations using accelerator mass spectrometry

Detection of Genotoxic Nucleosides, 8-Hydroxydeoxyguanosine, 8-Hydroxyguanosine and Free Base 8-Hydroxyguanine, in Fish Food Products

Contact Info

Product

Resources

About