DNA photoproducts released by repair in biological fluids as biomarkers of the genotoxicity of UV radiation

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Reynaud, Noémie | Belz, Laura | Béal, David | Bacqueville, Daniel | Duplan, Hélène | Genies, Camille | Questel, Emmanuel | Josse, Gwendal | Douki, Thierry

Edité par CCSD ; Springer Verlag -

International audience. UV-induced formation of photoproducts in DNA is a major initiating event of skin cancer. Consequently, many analytical tools have been developed for their quantification in DNA. In the present work, we extended our previous liquid chromatography-mass spectrometry method to the quantification of the short DNA fragments containing photoproducts that are released from cells by the repair machinery. We designed a robust protocol including a solid phase extraction step (SPE), an enzymatic treatment aimed at releasing individual photoproducts, and a liquid chromatography method combining on-line SPE and ultra-high performance liquid chromatography for optimal specificity and sensitivity. We also added relevant internal standards for a better accuracy. The method was validated for linearity, repeatability and reproducibility. The limits of detection and quantification were found to be in the fmol range. The proof of concept of the use of excreted DNA repair products as biomarkers of the genotoxicity of UV was obtained first in in vitro studies using cultured HaCat cells and ex-vivo on human skin explants. Further evidence were obtained from the detection of pyrimidine dimers in the urine of human volunteers collected after recreational exposure in summer.

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