INDUCTION and DISAPPEARANCE OF THYMINE DIMERS IN HUMAN SKIN EXPOSED TO UVB RADIATION: FLOW CYTOMETRIC MEASUREMENTS IN REPLICATING and NONREPLICATING EPIDERMAL CELLS |
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Authors: | Rob J W Berg Susan C de Bueger Kees Guikers Huib van Weelden Willem A van Vloten Jan C van der Leun Frank R de Gruul |
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Institution: | Department of Dermatology, University Hospital Utrecht, P.O. Box 85500, Utrecht, The Netherlands |
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Abstract: | We have earlier reported on determining UV-induced DNA damage in murine epidermal cell suspensions by flow cytometric analysis of the fluorescence from a fluorescein isothiocyanate-labeled antibody (H3) directed against thymine dimers (T>2-M-phase cells can further be distinguished from cell doublets by pulse-shape discrimination. Thus, T>(i.e. G0-G1, S or G2-M phases) can be determined after in vivo exposure of human skin to environmentally relevant UVB (280–315nm) doses. The method was applied to measure the decrease of T>0-G1 phase) and replicating cells (S phase or G2-M phase) from seven volunteers exposed to twice their minimal erythema dose. The reduction in the average T>0-G1 cells and 70% (ranging between 37% and 100%) for the S + G2-M cells. The difference was statistically highly significant. Determination of individual DNA repair capacities with this method can become a convenient diagnostic tool for patients with DNA repair disorders, or it may even be used to identify individuals with low repair proficiencies and increased risk of developing skin cancers. |
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