DNA damage induced by micro- and nanoparticles--interaction with FPG influences the detection of DNA oxidation in the comet assay.

2.50
Hdl Handle:
http://hdl.handle.net/10146/218671
Title:
DNA damage induced by micro- and nanoparticles--interaction with FPG influences the detection of DNA oxidation in the comet assay.
Authors:
Kain, J.; Karlsson, H. L.; Moller, L.
Abstract:
Reliable methods for evaluation of toxicity from particles, such as manufactured nanoparticles, are needed. One promising tool is the comet assay, often used to measure DNA breaks (strand breaks and alkali-labile sites) as well as oxidatively damaged DNA, the latter by addition of specific DNA repair enzymes such as formamidopyrimidine DNA glycosylase (FPG). The aim of this study was to investigate the use of the comet assay for analysis of DNA oxidation by a range of micro- and nanoparticles in the lung cell lines A549 and BEAS-2B and to test the hypothesis that nanoparticles present in the cells during the assay performance may interact with FPG. This was done by investigating the ability of micro- and nanoparticles (stainless steel, subway particles, MnO(2), Ag, CeO(2), Co(3)O(4), Fe(3)O(4), NiO and SiO(2)) to induce DNA breaks, oxidatively damaged DNA (FPG sites, dominantly 8-oxoguanine), intracellular production of reactive oxygen species (ROS) and non-cellular oxidation of the DNA base guanine, as well as by studying interactions of the particles and their released ions with FPG. Several particles caused DNA breaks, but low levels of FPG sites. The ability of FPG to detect DNA oxidation induced by a photosensitiser was however shown. An oxidative capacity of the particles was indicated by increased levels of intracellular ROS, and especially Ag and subway particles caused non-cellular oxidation of guanine. Incubation of FPG with the particles led to less FPG activity, particularly with nanoparticles of Ag but also with CeO(2), Co(3)O(4) and SiO(2). Further investigations of these particles revealed that for Ag, the decreased activity was mainly due to released Ag ions, whereas for CeO(2) and Co(3)O(4), FPG interactions were due to the particles. We conclude that measurement of oxidatively damaged DNA in cells exposed to nanoparticles may be underestimated in the comet assay due to interactions with FPG.
Citation:
Mutagenesis 2012, 27 (4):491-500
Journal:
Mutagenesis
Issue Date:
23-Mar-2012
URI:
http://hdl.handle.net/10146/218671
DOI:
10.1093/mutage/ges010
PubMed ID:
22447192
Additional Links:
http://mutage.oxfordjournals.org/content/early/2012/03/23/mutage.ges010.long
Type:
Article
Language:
en
ISSN:
1464-3804
Sponsors:
The authors are partners of ECNIS (Environmental Cancer Risk, Nutrition and Individual Susceptibility), an EU network of excellence operating within the European Union sixth framework program, priority 5: ‘Food Quality and Safety’ (contract no. 513943).
Appears in Collections:
Articles

Full metadata record

DC FieldValue Language
dc.contributor.authorKain, J.en_GB
dc.contributor.authorKarlsson, H. L.en_GB
dc.contributor.authorMoller, L.en_GB
dc.date.accessioned2012-04-17T08:45:05Z-
dc.date.available2012-04-17T08:45:05Z-
dc.date.issued2012-03-23-
dc.identifier.citationMutagenesis 2012, 27 (4):491-500en_GB
dc.identifier.issn1464-3804-
dc.identifier.pmid22447192-
dc.identifier.doi10.1093/mutage/ges010-
dc.identifier.urihttp://hdl.handle.net/10146/218671-
dc.description.abstractReliable methods for evaluation of toxicity from particles, such as manufactured nanoparticles, are needed. One promising tool is the comet assay, often used to measure DNA breaks (strand breaks and alkali-labile sites) as well as oxidatively damaged DNA, the latter by addition of specific DNA repair enzymes such as formamidopyrimidine DNA glycosylase (FPG). The aim of this study was to investigate the use of the comet assay for analysis of DNA oxidation by a range of micro- and nanoparticles in the lung cell lines A549 and BEAS-2B and to test the hypothesis that nanoparticles present in the cells during the assay performance may interact with FPG. This was done by investigating the ability of micro- and nanoparticles (stainless steel, subway particles, MnO(2), Ag, CeO(2), Co(3)O(4), Fe(3)O(4), NiO and SiO(2)) to induce DNA breaks, oxidatively damaged DNA (FPG sites, dominantly 8-oxoguanine), intracellular production of reactive oxygen species (ROS) and non-cellular oxidation of the DNA base guanine, as well as by studying interactions of the particles and their released ions with FPG. Several particles caused DNA breaks, but low levels of FPG sites. The ability of FPG to detect DNA oxidation induced by a photosensitiser was however shown. An oxidative capacity of the particles was indicated by increased levels of intracellular ROS, and especially Ag and subway particles caused non-cellular oxidation of guanine. Incubation of FPG with the particles led to less FPG activity, particularly with nanoparticles of Ag but also with CeO(2), Co(3)O(4) and SiO(2). Further investigations of these particles revealed that for Ag, the decreased activity was mainly due to released Ag ions, whereas for CeO(2) and Co(3)O(4), FPG interactions were due to the particles. We conclude that measurement of oxidatively damaged DNA in cells exposed to nanoparticles may be underestimated in the comet assay due to interactions with FPG.en_GB
dc.description.sponsorshipThe authors are partners of ECNIS (Environmental Cancer Risk, Nutrition and Individual Susceptibility), an EU network of excellence operating within the European Union sixth framework program, priority 5: ‘Food Quality and Safety’ (contract no. 513943).en_GB
dc.languageENG-
dc.language.isoenen
dc.relation.urlhttp://mutage.oxfordjournals.org/content/early/2012/03/23/mutage.ges010.longen_GB
dc.rightsArchived with thanks to Mutagenesisen_GB
dc.subjectNanoparticlesen_GB
dc.subjectToxicityen_GB
dc.subjectDNA damageen_GB
dc.subjectDNA oxidationen_GB
dc.subjectDNA repair enzymesen_GB
dc.subjectCell cultureen_GB
dc.subjectLung cell linesen_GB
dc.subjectComet assayen_GB
dc.subject.meshBronchien
dc.subject.meshCells, Cultureden
dc.subject.meshChromatography, High Pressure Liquiden
dc.subject.meshComet Assayen
dc.subject.meshDNAen
dc.subject.meshDNA Damageen
dc.subject.meshDNA-Formamidopyrimidine Glycosylaseen
dc.subject.meshGuanineen
dc.subject.meshHumansen
dc.subject.meshMetal Nanoparticlesen
dc.subject.meshOxidation-Reductionen
dc.subject.meshOxidative Stressen
dc.subject.meshPulmonary Alveolien
dc.subject.meshReactive Oxygen Species-
dc.titleDNA damage induced by micro- and nanoparticles--interaction with FPG influences the detection of DNA oxidation in the comet assay.en
dc.typeArticleen
dc.identifier.journalMutagenesisen_GB

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