Functional evaluation of DNA repair in human biopsies and their relation to other cellular biomarkers
Thousands of DNA lesions are estimated to occur in each cell every day and almost all are recognized and repaired. DNA repair is an essential system that prevents accumulation of mutations which can lead to serious cellular malfunctions. Phenotypic evaluation of DNA repair activity of individuals is...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2014-05-01
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| Series: | Frontiers in Genetics |
| Subjects: | |
| Online Access: | http://journal.frontiersin.org/Journal/10.3389/fgene.2014.00116/full |
| _version_ | 1818612012693848064 |
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| author | Jana eSlyskova Sabine eLangie Andrew eCollins Pavel eVodicka |
| author_facet | Jana eSlyskova Sabine eLangie Andrew eCollins Pavel eVodicka |
| author_sort | Jana eSlyskova |
| collection | DOAJ |
| description | Thousands of DNA lesions are estimated to occur in each cell every day and almost all are recognized and repaired. DNA repair is an essential system that prevents accumulation of mutations which can lead to serious cellular malfunctions. Phenotypic evaluation of DNA repair activity of individuals is a relatively new approach. Methods to assess base and nucleotide excision repair pathways (BER and NER) in peripheral blood cells based on modified comet assay protocols have been widely applied in human epidemiological studies. These provided some interesting observations of individual DNA repair activity being suppressed among cancer patients. However, extension of these results to cancer target tissues requires a different approach. Here we describe the evaluation of BER and NER activities in extracts from deep-frozen colon biopsies using an upgraded version of the in vitro comet-based DNA repair assay in which twelve reactions on one microscope slide can be performed. The aim of this report is to provide a detailed, easy-to-follow protocol together with results of optimization experiments. Additionally, results obtained by functional assays were analysed in the context of other cellular biomarkers, namely single nucleotide polymorphisms and gene expressions. We have shown that measuring DNA repair activity is not easily replaceable by genomic or transcriptomic approaches, but should be applied with the latter techniques in a complementary manner. The ability to measure DNA repair directly in cancer target tissues might finally answer questions about the tissue-specificity of DNA repair processes and their real involvement in the process of carcinogenesis. |
| first_indexed | 2024-12-16T15:39:27Z |
| format | Article |
| id | doaj.art-38c7bf05ad12484f9a1881368be4c979 |
| institution | Directory Open Access Journal |
| issn | 1664-8021 |
| language | English |
| last_indexed | 2024-12-16T15:39:27Z |
| publishDate | 2014-05-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Genetics |
| spelling | doaj.art-38c7bf05ad12484f9a1881368be4c9792022-12-21T22:26:04ZengFrontiers Media S.A.Frontiers in Genetics1664-80212014-05-01510.3389/fgene.2014.0011691228Functional evaluation of DNA repair in human biopsies and their relation to other cellular biomarkersJana eSlyskova0Sabine eLangie1Andrew eCollins2Pavel eVodicka3Institute of Experimental Medicine, Academy of Sciences of the Czech RepublicFlemish Institute for Technological Research (VITO)University of OsloInstitute of Experimental Medicine, Academy of Sciences of the Czech RepublicThousands of DNA lesions are estimated to occur in each cell every day and almost all are recognized and repaired. DNA repair is an essential system that prevents accumulation of mutations which can lead to serious cellular malfunctions. Phenotypic evaluation of DNA repair activity of individuals is a relatively new approach. Methods to assess base and nucleotide excision repair pathways (BER and NER) in peripheral blood cells based on modified comet assay protocols have been widely applied in human epidemiological studies. These provided some interesting observations of individual DNA repair activity being suppressed among cancer patients. However, extension of these results to cancer target tissues requires a different approach. Here we describe the evaluation of BER and NER activities in extracts from deep-frozen colon biopsies using an upgraded version of the in vitro comet-based DNA repair assay in which twelve reactions on one microscope slide can be performed. The aim of this report is to provide a detailed, easy-to-follow protocol together with results of optimization experiments. Additionally, results obtained by functional assays were analysed in the context of other cellular biomarkers, namely single nucleotide polymorphisms and gene expressions. We have shown that measuring DNA repair activity is not easily replaceable by genomic or transcriptomic approaches, but should be applied with the latter techniques in a complementary manner. The ability to measure DNA repair directly in cancer target tissues might finally answer questions about the tissue-specificity of DNA repair processes and their real involvement in the process of carcinogenesis.http://journal.frontiersin.org/Journal/10.3389/fgene.2014.00116/fullnucleotide excision repairBase excision repairHuman solid tissueMethodological reportin vitro comet-based DNA repair assay |
| spellingShingle | Jana eSlyskova Sabine eLangie Andrew eCollins Pavel eVodicka Functional evaluation of DNA repair in human biopsies and their relation to other cellular biomarkers Frontiers in Genetics nucleotide excision repair Base excision repair Human solid tissue Methodological report in vitro comet-based DNA repair assay |
| title | Functional evaluation of DNA repair in human biopsies and their relation to other cellular biomarkers |
| title_full | Functional evaluation of DNA repair in human biopsies and their relation to other cellular biomarkers |
| title_fullStr | Functional evaluation of DNA repair in human biopsies and their relation to other cellular biomarkers |
| title_full_unstemmed | Functional evaluation of DNA repair in human biopsies and their relation to other cellular biomarkers |
| title_short | Functional evaluation of DNA repair in human biopsies and their relation to other cellular biomarkers |
| title_sort | functional evaluation of dna repair in human biopsies and their relation to other cellular biomarkers |
| topic | nucleotide excision repair Base excision repair Human solid tissue Methodological report in vitro comet-based DNA repair assay |
| url | http://journal.frontiersin.org/Journal/10.3389/fgene.2014.00116/full |
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