The relationship between cellular radiosensitivity and radiation-induced DNA damage measured by the comet assay.
The relationship between deoxyribonucleic acid (DNA) damage and the cell death induced by gamma-irradiation was examined in three kinds of cells, Chinese hamster ovary fibroblast CHO-K1, human melanoma HMV-II and mouse leukemia L5178Y. Cell survival was determined by a clonogenic assay. The induction and rejoining of DNA strand breaks induced by radiation were measured by the alkaline and neutral comet assays. L5178Y cells were the most radiosensitive, while CHO-K1 cells and HMV-II cells were radioresistant. There was an inverse relationship between the survival fraction at 2 Gy (SF2) and the yield of initial DNA strand breaks per unit dose under the alkaline condition for the comet assay, and also a relationship between SF2 and the residual DNA strand breaks (for 4 hr after irradiation) under the neutral condition for the comet assay, the latter being generally considered to be relative to cellular radiosensitivity. In the present analysis, it was considered that the alkaline condition for the comet assay was optimal for evaluating the initial DNA strand breaks, while the neutral condition was optimal for evaluating the residual DNA strand breaks. Since the comet assay is simpler and more rapid than other methods for detecting radiation-induced DNA damage, this assay appears to be a useful predictive assay for evaluating cellular clonogenic radiosensitivity of tumor cells.
Wada
Seiichi
S
Biotechnology Laboratory, JAERI-Takasaki, Watanuki-machi, Gunma, Japan.
Kurahayashi
Hidemitsu
H
Kobayashi
Yasuhiko
Y
Funayama
Tomoo
T
Yamamoto
Kazuo
K
Natsuhori
Masahiro
M
Ito
Nobuhiko
N
D000818:Animals; D016466:CHO Cells; D016923:Cell Death / Q000235:genetics / Q000528:radiation effects*; D002470:Cell Survival / Q000235:genetics / Q000528:radiation effects*; D020552:Comet Assay / Q000662:veterinary*; D006224:Cricetinae; D004249:DNA Damage; D004307:Dose-Response Relationship, Radiation; D005720:Gamma Rays; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007938:Leukemia; D016014:Linear Models; D008545:Melanoma; D051379:Mice; D014407:Tumor Cells, Cultured