Comet assay to measure DNA repair: approach and applications

Azqueta, Amaya; Slyšková, Jana; Langie, Sabine A. S.; Gaivão, Isabel; Collins, Andrew

doi:10.3389/fgene.2014.00288

Cited by 146 publications

(114 citation statements)

References 60 publications

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“…Taking into account that the majority of primary DNA lesions detectable using the alkaline comet assay are potentially repairable, our results mostly indicate the acute effects caused by the tested compounds, and also suggest the differences in their ability to destabilise the integrity of DNA in kidney cells of mice. It is noteworthy to mention here that the alkaline comet assay we used allows a sensitive detection of different forms of primary DNA damage in single cells but the most important among them are DNA strand breaks and alkali-labile sites (8,(21)(22)(23).…”

Section: Discussionmentioning

confidence: 99%

Sevoflurane and isoflurane genotoxicity in kidney cells of mice

Brozović

Oršolić

Rozgaj

et al. 2017

Archives of Industrial Hygiene and Toxicology

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The aim of this study was to evaluate the DNA damage and repair in kidney cells of Swiss albino mice after repeated exposure to sevoflurane and isoflurane and compare their detrimental effects. We used the alkaline comet assay to establish the genetic damage and measured three parameters: tail length, tail moment, and tail intensity of comets. These parameters were measured immediately after exposure to the above mentioned inhalation anaesthetics, two hours, six hours, and 24 hours later and were compared with the control group. Mean values of all three parameters were significantly higher in experimental groups compared to the control group. DNA damage in kidney cells of mice exposed to sevoflurane increased continuously before it reached its peak 24 hours after exposure. Isoflurane induced the highest DNA damage two hours after exposure. Levels of DNA damage recorded 24 h after cessation of exposure to both tested compounds suggest that sevoflurane was slightly more genotoxic than isoflurane to kidney cells of mice. According to these results, the currently used volatile anaesthetics sevoflurane and isoflurane are able to damage DNA in kidney cells of mice. Such findings suggest a possibility for similar outcomes in humans and that fact must be taken into account in everyday clinical practice.

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Section: Discussionmentioning

confidence: 99%

Sevoflurane and isoflurane genotoxicity in kidney cells of mice

Brozović

Oršolić

Rozgaj

et al. 2017

Archives of Industrial Hygiene and Toxicology

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“…To estimate the levels of primary DNA damage in hydroquinone-treated lymphocytes, we used an alkaline version of the comet assay, which detects the widest range of lesions (56)(57)(58). First literature reports regarding hydroquinone effects detected by the comet assay in peripheral blood lymphocytes treated in vitro originated from Andreoli et al (59) who found that hydroquinone, along with two other benzene metabolites -benzoquinone and benzenetriol -exerted a significant genotoxic effect on resting lymphocytes, with the lowest effective concentrations below 1 µg mL -1 .…”

Section: Discussionmentioning

confidence: 99%

In vitro assessment of the cytotoxic, DNA damaging, and cytogenetic effects of hydroquinone in human peripheral blood lymphocytes

Jurica

Karačonji

Benković

et al. 2017

Archives of Industrial Hygiene and Toxicology

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This study investigated the mechanisms of hydroquinone toxicity and assessed the relationships between its cytotoxic, genotoxic, and cytogenetic effects tested at 8, 140, and 280 µg mL -1 in human peripheral blood lymphocytes exposed for 24 h. The outcomes of the treatments were evaluated using the apoptosis/necrosis assay, the alkaline comet assay, and the cytokinesis-block micronucleus (CBMN) cytome assay. The tested hydroquinone concentrations produced relatively weak cytotoxicity in resting lymphocytes, which mostly died via apoptosis. Hydroquinone's marked genotoxic effects were detected using the alkaline comet assay. Significantly decreased values of all comet parameters compared to controls indicated specific mechanisms of hydroquinone-DNA interactions. Our results suggest that the two higher hydroquinone concentrations possibly led to cross-linking and adduct formation. Increased levels of DNA breakage measured following exposure to the lowest concentration suggested mechanisms related to oxidative stress and inhibition of topoisomerase II. At 8 µg mL -1 , hydroquinone did not significantly affect MN formation. At 140 and 280 µg mL -1 , it completely blocked lymphocyte division. The two latter concentrations also led to erythrocyte stabilization and prevented their lysis. At least two facts contribute to this study's relevance: (I) this is the first study that quantifies the degree of reduction in total comet area measured in lymphocyte DNA after hydroquinone treatment, (II) it is also the first one on a lymphocyte model that adopted the "cytome" protocol in an MN assay and found that lymphocytes exposure even to low hydroquinone concentration resulted in a significant increase of nuclear bud frequency. Considering the limitations of the lymphocyte model, which does not possess intrinsic metabolic activation, in order to unequivocally prove the obtained results further studies using other appropriate cell lines are advised.

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“…However, in the case of comet assay, the positive results could be attributed to the fact that the transient promutagenic DNA changes, e.g. SSBs, might be quickly repaired (Azqueta et al 2014). Hence, these photogenotoxic findings only have an informative value, in contrast to gene and chromosome mutations.…”

Section: Discussionmentioning

confidence: 89%

Photoactivated hypericin is not genotoxic

Feruszova¹,

Imreova²,

Bodnarova³

et al. 2016

gpb

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Abstract. The study was designed to test the potential photogenotoxicity of hypericin (HYP) at three different levels: primary DNA damages, gene mutations and chromosome aberrations. Primary genetic changes were detected using the comet assay. The potential mutagenic activity of HYP was assessed using the Ames/Salmonella typhimurium assay. Finally, the ability of photoactivated HYP to induce chromosome aberrations was evaluated by the in vitro mammalian chromosome aberration test and compared to that of non-photoactivated HYP. The results have shown that photoactivated HYP can only induce primary DNA damages (single-strand DNA breaks), acting in a dose-response manner. This activity depended both on HYP concentrations and an intensity of the light energy needed for its photoactivation. However, mutagenic effect of photoactivated HYP evaluated in the Ames assay using three bacterial strains S. typhimurium (TA97, TA98 and TA100) was not confirmed. Moreover, photoactivated HYP in the range of concentrations (0.005-0.01 µg/ml) was not found to be clastogenic against HepG2 cells. Our findings from both the Ames assay and the chromosome aberrations test provide evidence that photoactivated HYP is not genotoxic, which might be of great importance mainly in terms of its use in the photodynamic therapy.

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Comet assay to measure DNA repair: approach and applications

Cited by 146 publications

References 60 publications

Sevoflurane and isoflurane genotoxicity in kidney cells of mice

Sevoflurane and isoflurane genotoxicity in kidney cells of mice

In vitro assessment of the cytotoxic, DNA damaging, and cytogenetic effects of hydroquinone in human peripheral blood lymphocytes

Photoactivated hypericin is not genotoxic

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