Comparative study with the alkaline Comet assay and the chromosome aberration test

Hartmann, Andreas; Plappert, U.; Poetter, Franziska; Suter, Willi

doi:10.1016/s1383-5718(03)00035-4

Cited by 63 publications

(20 citation statements)

References 27 publications

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“…Singlecell gel electrophoresis (comet assay) provides a very sensitive method for detecting DNA damage at the single cell level (Collins et al, 1995;Brendler-Schwaab et al, 2005). The alkaline comet assay is increasingly used in genotoxicity testing in vitro and is also becoming an important tool for evaluating the genotoxic potential of compounds in vivo (Hartmann et al, 2003;Brendler-Schwaab et al, 2005). Among the advantages of the comet assay are its sensitivity for detecting low levels of DNA damage, the need for only a small number of cells per sample, and its applicability to any kind of eukaryotic organism and cell type (Silva et al, 2000;Brendler-Schwaab et al, 2005).…”

Section: Introductionmentioning

confidence: 99%

Antioxidant effect of haptoglobin phenotypes against DNA damage induced by hydrogen peroxide in human leukocytes

Moreira

Miranda-Vilela²,

Silva³

et al. 2009

Genet. Mol. Res.

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ABSTRACT. Human haptoglobin is classified into three major phenotypes: Hp1-1, Hp2-1 and Hp2-2; there are two autosomal alleles Hp . Haptoglobin acts as an antioxidant, preventing hemoglobin-driven oxidative damage. We used the comet assay to examine oxidative damage to DNA induced by hydrogen peroxide in human leukocytes; we also looked for differences in the antioxidant capacity of haptoglobin subtypes. Haptoglobin genotypes were determined through allele-specific polymerase chain reaction, visualized on a polyacrylamide gel. The Hp1-1 genotype had the least DNA damage; this indicates that Hp alleles differ in their protective effects against oxidative damage. Among Hp *1 alleles, Hp *1F was the most protective.

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Section: Introductionmentioning

confidence: 99%

Antioxidant effect of haptoglobin phenotypes against DNA damage induced by hydrogen peroxide in human leukocytes

Moreira

Miranda-Vilela²,

Silva³

et al. 2009

Genet. Mol. Res.

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“…Compared to other tests for genotoxic damage, the principal advantages of the comet assay include its demonstrated sensitivity for detecting DNA damage at a very low degree, the requirement for a small number of cells per sample, its flexibility, low cost, and the relatively short time needed to complete a study. [43][44][45] The data collected in the present study suggest that the comet assay could contribute to treatment decisions by considering the patient's basal DNA damage before therapy. At present, therapy decisions are largely based on prognostic factors, such as the presence of node metastases, among others.…”

Section: A U T H O R P R O O Fmentioning

confidence: 80%

Relationship between Genotoxic Effects of Breast Cancer Treatments and Patient Basal DNA Integrity

Ceballos

Funes

Massa

et al. 2014

J Environ Pathol Toxicol Oncol

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Radiotherapy and chemotherapy cause genotoxic side effects that are highly variable among patients. In this study, we evaluated DNA integrity using the comet assay in peripheral blood lymphocytes from breast cancer patients before ("pre-treatment patients"; n = 47) and after ("post-treatment patients"; n = 24) radiotherapy and/or chemotherapy treatment and from healthy donors (n = 15). Comet evaluation was made by visual (types 0-4) and digital (percentage of DNA remaining in the comet head = % head DNA) analysis. The association between the level of DNA damage and cancer prognostic factors was assessed. The treatments caused a significant increase in DNA damage registered by both visual (p < 0.001) and digital (p < 0.001) analyses. No significant associations between the level of DNA damage in pre-treatment patients and cancer prognostic factors were found. A significant correlation between the comet results from each patient before and after treatment (r = 0.64, p = 0.001) was observed. The % head DNA in post-treatment samples from patients with a high level of DNA damage before treatment (30.3 ± 3.1%, p < 0.01) was lower than in post-treatment samples from patients with a low-to-medium level of DNA damage before therapy (49.2 ± 4.4%). These results support the usefulness of the comet assay as a sensitive technique to evaluate basal DNA status and DNA damage caused by cancer treatments. The comet assay could contribute to treatment decisions, especially by taking into account the patient's basal DNA damage before therapy.

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“…This test allows the detection of DNA damage such as single and double-strand breaks and alkali labile lesions in individual cells after acute and/or chronic exposure to a genotoxic agent (Tice et al, 2000). According to Hartmann et al (2003) the chromosome aberration assay can be used to validate the results of the Comet assay.…”

Section: Discussionmentioning

confidence: 99%

“…It provides a straightforward visual method for assessing DNA damage quantitatively in single cells and is established as a valuable tool in fundamental DNA damage and repair studies (Visvardis et al, 2000). The chromosome aberration assay is also a powerful classical cytogenetic tool for genotoxicity testing and can be used as a validation test for Comet assay results (Hartmann et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

Genotoxic and antigenotoxic effects of Fucus vesiculosus extract on cultured human lymphocytes using the chromosome aberration and Comet assays

2007

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The brown seaweed Fucus vesiculosus (Fucales, Fucaceae) was screened for its protective activity using doxorubicin-induced DNA damage in human lymphocytes. In this study, we assessed the genotoxic and antigenotoxic potential of three different concentrations (0.25, 0.5 and 1.0 mg mL -1 ) of F. vesiculosus aqueous extract using the chromosome aberration and Comet assays. Treatment of human lymphocyte cultures with 0.25, 0.5 and 1.0 mg mL -1 F. vesiculosus aqueous extract had no effect on the chromosome aberration frequency or on the extent of DNA damage detected by the Comet assay. The antigenotoxic effects of the extract were tested in human lymphocyte cultures treated with 15 μg mL -1 of doxorubicin, either alone or combined with the different concentrations of the extract, which was added to the cultures before, simultaneously with or after the doxorubicin. Only when lymphocytes were pre-treated with extract there was a reduction in doxorubicin-induced chromosome aberrations and DNA damage as detected by the Comet assay. These results demonstrate that F. vesiculosus aqueous extract is not genotoxic in cultured human lymphocytes and indicate that when added to lymphocyte cultures before doxorubicin it has antigenotoxic activity against doxorubicin-induced DNA damage.

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Comparative study with the alkaline Comet assay and the chromosome aberration test

Cited by 63 publications

References 27 publications

Antioxidant effect of haptoglobin phenotypes against DNA damage induced by hydrogen peroxide in human leukocytes

Antioxidant effect of haptoglobin phenotypes against DNA damage induced by hydrogen peroxide in human leukocytes

Relationship between Genotoxic Effects of Breast Cancer Treatments and Patient Basal DNA Integrity

Genotoxic and antigenotoxic effects of Fucus vesiculosus extract on cultured human lymphocytes using the chromosome aberration and Comet assays

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