Radiation-induced comet-formation in human skin fibroblasts from radiotherapy patients with different normal tissue reactions.
BACKGROUND
In clinical radiotherapy most patients tolerate the applied dosage with no or moderate side effects. However, 5 to 10% of all individuals show increased acute and/or late reactions. In-vitro test systems are investigated for their suitability for predictive purposes. This paper attempts a correlation between the induction and repair of DNA damage measured in the comet assay and the clinical observed reaction in order to evaluate the suitability of the comet assay for prediction of radiation sensitivity.


METHODS
Skin fibroblasts of 30 patients with average tissue reactions or acute and/or late increased side effects and cell lines of 4 individuals carrying the heritable disease ataxia telangiectasia (AT) were irradiated in vitro. The induction and repair of DNA damage was measured at different time points after irradiation in the comet assay (single cell gel electrophoresis). These results were compared to the acute and late clinical reactions classified according to the RTOG grading system.


RESULTS
The radiation induced DNA damage decreased over time reflecting DNA repair. Cells of the AT individuals showed an elevated damage induction and a reduced repair capacity compared to patients with average tissue reactions. Fibroblasts of patients with increased acute and late side effects exhibited slower DNA repair. In addition to the known lack of cell cycle control, our results indicate that AT cells show reduced DNA repair capacity.


CONCLUSIONS
The comet assay seems to be able to detect some types of increased individual radiation sensitivity. In contrast to other predictive in-vitro tests, the comet assay needs less time and fewer cells, which would be useful in a clinical setting.
Oppitz
U
U
Klinik und Poliklinik für Strahlentherapie, Universität Würzburg, Germany. oppitz@strahlentherapie.uni-wuerzburg.de
Denzinger
S
S
Nachtrab
U
U
Flentje
M
M
Stopper
H
H
D001260:Ataxia Telangiectasia / Q000235:genetics / Q000473:pathology; D002455:Cell Division / Q000528:radiation effects; D004249:DNA Damage; D004260:DNA Repair / Q000528:radiation effects; D004307:Dose-Response Relationship, Radiation; D004587:Electrophoresis, Agar Gel; D005260:Female; D005347:Fibroblasts / Q000473:pathology / Q000528:radiation effects; D006801:Humans; D008297:Male; D009369:Neoplasms / Q000532:radiotherapy*; D011855:Radiodermatitis / Q000473:pathology*; D012867:Skin / Q000473:pathology / Q000528:radiation effects*