Use of the alkaline comet assay to detect DNA repair deficiencies in human fibroblasts exposed to UVC, UVB, UVA and gamma-rays.
The alkaline single cell gel electrophoresis (comet) assay applied to human fibroblasts allowed us to analyze the response to components of the solar spectrum (UVB and UVA) in comparison with the well-established response to UVC and gamma-rays. DNA strand breaks related to nucleotide excision repair of DNA photoproducts were produced 1 h after exposure to UVB or UVC in the normal cell line but not in the repair deficient XPD and TTD-2 cell lines. In contrast, the immediate production of DNA strand breaks observed in all cell lines after exposure to UVA or gamma-rays was followed by restitution of high molecular weight DNA upon post-exposure incubation. These results imply that (1) fibroblasts as well as lymphocytes can be analysed by the comet assay and (2) the comet assay clearly distinguishes cellular nucleotide excision repair capacity without the use of inhibitors of DNA synthesis.
Alapetite
C
C
URA 1292 du CNRS, Institut Curie-Section de Recherche, Institut Curie, Paris, France.
Wachter
T
T
Sage
E
E
Moustacchi
E
E
D002460:Cell Line; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification* / Q000528:radiation effects*; D004249:DNA Damage; D004260:DNA Repair; D004307:Dose-Response Relationship, Radiation; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D005347:Fibroblasts / Q000528:radiation effects*; D005720:Gamma Rays / Q000009:adverse effects; D006801:Humans; D009152:Mutagenicity Tests; D009381:Neoplasms, Radiation-Induced / Q000209:etiology; D010777:Photochemistry; D012878:Skin Neoplasms / Q000209:etiology; D014466:Ultraviolet Rays / Q000009:adverse effects
D004247:DNA