title: Nick translation detection in situ of cellular DNA strand break induced by radiation.
abstract: DNA strand break in HeLa cells induced by radiation was detected using the in situ nick translation method. The cells were exposed to radiation of 3, 6, 12, 18, and 24 Gy in Lab-Tek tissue culture chamber/slides and were fixed with ethanol/acetic acid on the slide glass. The break sites in DNA were translated artificially in the presence of Escherichia coli DNA polymerase I and [3H]-labeled dTTP. Autoradiographic observation was made of the level of break sites in the DNA. The DNA strand break appeared even with a 3 Gy exposure, increased 8.6 times at 24 Gy compared with the control cells, and this level correlated reciprocally to change in cell viability. This nick translation method provides a rapid in situ assay for determining radiation-induced DNA damage of cultured cells, in a semi-quantitative manner.
authors.affiliation: Cancer Center of Kyushu University Hospital, Faculty of Medicine, Kyushu University, Fukuoka, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Maehara
authors.forename: H
authors.initials: H
authors.lastname: Anai
authors.forename: T
authors.initials: T
authors.lastname: Kusumoto
authors.forename: Y
authors.initials: Y
authors.lastname: Sakaguchi
authors.forename: K
authors.initials: K
authors.lastname: Sugimachi
mesh_terms: D000818:Animals; D004247:DNA / Q000528:radiation effects; D004249:DNA Damage; D004307:Dose-Response Relationship, Radiation; D005821:Genetic Techniques; D006367:HeLa Cells / Q000528:radiation effects*; D006801:Humans; D014176:Protein Biosynthesis
chemical_list: D004247:DNA