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  • Thousands of DNA lesions 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 12 reactions on one microscope slide can be performed. Additionally, results obtained by functional assays were analyzed in the context of other cellular biomarkers, namely single nucleotide polymorphisms (SNPs) 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.
  • Thousands of DNA lesions 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 12 reactions on one microscope slide can be performed. Additionally, results obtained by functional assays were analyzed in the context of other cellular biomarkers, namely single nucleotide polymorphisms (SNPs) 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. (en)
Title
  • Functional evaluation of DNA repair in human biopsies and their relation to other cellular biomarkers
  • Functional evaluation of DNA repair in human biopsies and their relation to other cellular biomarkers (en)
skos:prefLabel
  • Functional evaluation of DNA repair in human biopsies and their relation to other cellular biomarkers
  • Functional evaluation of DNA repair in human biopsies and their relation to other cellular biomarkers (en)
skos:notation
  • RIV/68378041:_____/14:00432504!RIV15-GA0-68378041
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  • I, P(GAP304/12/1585)
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  • 5
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  • 17657
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  • RIV/68378041:_____/14:00432504
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  • base excision repair; nucleotide excision repair; human solid tissue (en)
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  • CH - Švýcarská konfederace
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  • [20D858364645]
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  • Frontiers in genetics
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  • 116
http://linked.open...iv/tvurceVysledku
  • Vodička, Pavel
  • Slyšková, Jana
  • Collins, A. R.
  • Langie, S. A. S.
issn
  • 1664-8021
number of pages
http://bibframe.org/vocab/doi
  • 10.3389/fgene.2014.00116
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