Nick translation detection in situ of cellular DNA strand break induced by radiation

Yoshihiko Maehara, H. Anai, T. Kusumoto, Y. Sakaguchi, K. Sugimachi

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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.

Original languageEnglish
Pages (from-to)7-10
Number of pages4
JournalAmerican Journal of Pathology
Volume134
Issue number1
Publication statusPublished - Jan 1 1989

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All Science Journal Classification (ASJC) codes

  • Pathology and Forensic Medicine

Cite this

Maehara, Y., Anai, H., Kusumoto, T., Sakaguchi, Y., & Sugimachi, K. (1989). Nick translation detection in situ of cellular DNA strand break induced by radiation. American Journal of Pathology, 134(1), 7-10.