Detection of carcinogen-induced DNA breaks by nick translation in permeable cells

Kiyoshi Nose, Hiroshi Okamoto

Research output: Contribution to journalArticlepeer-review

42 Citations (Scopus)

Abstract

A nick-translation reaction with E. coli DNA polymerase I (pol. I) was used to detect in situ DNA breaks produced by chemical carcinogens. Normal human fibroblasts treated with N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) in various doses were permeabilized with lysolecithin, and were nick translated in the presence of [3H]dCTP and pol. I. The radioactivity incorporated increased with MNNG concentration, and was directly proportional to the poly(ADP-ribose) synthetase activity. Other DNA-damaging agents such as bleomycin or 4-nitroquinoline 1-oxide also caused the nick translation rate to increase. When MNNG-treated cells were cultured in fresh medium containing no MNNG, the increase in the rate of nick translation in permeable cells became less and this decrease was abolished by addition of aphidicolin or cytosine arabinoside. The nick translation method described here may be a useful means for estimating intrinsic DNA breaks in cells treated with carcinogens.

Original languageEnglish
Pages (from-to)383-389
Number of pages7
JournalBiochemical and biophysical research communications
Volume111
Issue number2
DOIs
Publication statusPublished - 1983 Mar 16
Externally publishedYes

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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