Oxidative DNA damage in cultured fibroblasts from patients with hereditary glutathione synthetase deficiency |
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Authors: | Nygren Jonas Ristoff Ellinor Carlsson Katarina Möller Lennart Larsson Agne |
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Institution: |
a Department of Industrial Hygiene and Toxicology, Laboratory of Molecular and Cellular Toxicology, Finnish Institute of Occupational Health, Helsinki, Finland
b Department of Pediatrics, Karolinska Institutet, Karolinska University Hospital Huddinge, Stockholm, Sweden
c Department of Biosciences at Novum, Karolinska Institutet, Stockholm, Sweden |
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Abstract: | The SH compound glutathione (GSH) is involved in several fundamental functions in the cell, including protection against reactive oxygen species (ROS). Here, we studied the effect on oxidative DNA damage in cultured skin fibroblasts from patients with hereditary GSH synthetase deficiency. Our hypothesis was that GSH-deficient cells are more prone to DNA damage than control cells. Single cell gel electrophoresis (the comet assay) in combination with the formamidopyrimidine DNA glycosylase enzyme, which recognizes oxidative base modifications, was used on cultured fibroblasts from 11 patients with GSH synthetase deficiency and five control subjects. Contrary to this hypothesis, we found no significant difference in background levels of DNA damage between cells from patients and control subjects. To study the induction of oxidative DNA damage without simultaneous DNA repair, the cells were γ-irradiated on ice and DNA single-strand breaks measured. The patient and control cells were equally sensitive to induction of single strand breaks by γ-irradiation. Therefore, factors other than GSH protect DNA from oxidative damage. However, cells with a high background level of oxidative DNA damage were found to be more sensitive to ionizing radiation. This suggests that differences in background levels of oxidative DNA damage may depend on the cells' intrinsic protection against induction of oxidative damage. |
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Keywords: | Glutathione glutathione synthetase glutathione synthetase deficiency comet assay DNA damage oxidative stress |
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