In situ nick translation method reveals DNA strand scission in HeLa cells following heat treatment

Hideaki Anai, Yoshihiko Maehara, Keizo Sugimachi

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

DNA strand break in HeLa cells induced by heat was detected using the in situ nick translation method. The cells were incubated at 43°C for various times (15, 30, 45, 60, 90 or 120 min) 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. The level of break sites in the DNA was visualized by autoradiographic observation of the grains. The DNA strand break appeared as early as 15 min, increased to 10.3-fold at 45 min of 43°C treatment and this level related reciprocally to clonogenicity of the cell. The nick translation method thus provides a rapid in situ assay for determining heat-induced DNA damage of cultured cells, in a semi-quantitative manner.

Original languageEnglish
Pages (from-to)33-38
Number of pages6
JournalCancer Letters
Volume40
Issue number1
DOIs
Publication statusPublished - May 1988

Fingerprint

DNA Breaks
HeLa Cells
Hot Temperature
DNA Polymerase I
DNA
Acetic Acid
DNA Damage
Glass
Cultured Cells
Ethanol
Observation
Escherichia coli
thymidine 5'-triphosphate

All Science Journal Classification (ASJC) codes

  • Oncology
  • Cancer Research

Cite this

In situ nick translation method reveals DNA strand scission in HeLa cells following heat treatment. / Anai, Hideaki; Maehara, Yoshihiko; Sugimachi, Keizo.

In: Cancer Letters, Vol. 40, No. 1, 05.1988, p. 33-38.

Research output: Contribution to journalArticle

Anai, Hideaki ; Maehara, Yoshihiko ; Sugimachi, Keizo. / In situ nick translation method reveals DNA strand scission in HeLa cells following heat treatment. In: Cancer Letters. 1988 ; Vol. 40, No. 1. pp. 33-38.
@article{e38b43908ca3416c9594729d24680392,
title = "In situ nick translation method reveals DNA strand scission in HeLa cells following heat treatment",
abstract = "DNA strand break in HeLa cells induced by heat was detected using the in situ nick translation method. The cells were incubated at 43°C for various times (15, 30, 45, 60, 90 or 120 min) 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. The level of break sites in the DNA was visualized by autoradiographic observation of the grains. The DNA strand break appeared as early as 15 min, increased to 10.3-fold at 45 min of 43°C treatment and this level related reciprocally to clonogenicity of the cell. The nick translation method thus provides a rapid in situ assay for determining heat-induced DNA damage of cultured cells, in a semi-quantitative manner.",
author = "Hideaki Anai and Yoshihiko Maehara and Keizo Sugimachi",
year = "1988",
month = "5",
doi = "10.1016/0304-3835(88)90259-5",
language = "English",
volume = "40",
pages = "33--38",
journal = "Cancer Letters",
issn = "0304-3835",
publisher = "Elsevier Ireland Ltd",
number = "1",

}

TY - JOUR

T1 - In situ nick translation method reveals DNA strand scission in HeLa cells following heat treatment

AU - Anai, Hideaki

AU - Maehara, Yoshihiko

AU - Sugimachi, Keizo

PY - 1988/5

Y1 - 1988/5

N2 - DNA strand break in HeLa cells induced by heat was detected using the in situ nick translation method. The cells were incubated at 43°C for various times (15, 30, 45, 60, 90 or 120 min) 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. The level of break sites in the DNA was visualized by autoradiographic observation of the grains. The DNA strand break appeared as early as 15 min, increased to 10.3-fold at 45 min of 43°C treatment and this level related reciprocally to clonogenicity of the cell. The nick translation method thus provides a rapid in situ assay for determining heat-induced DNA damage of cultured cells, in a semi-quantitative manner.

AB - DNA strand break in HeLa cells induced by heat was detected using the in situ nick translation method. The cells were incubated at 43°C for various times (15, 30, 45, 60, 90 or 120 min) 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. The level of break sites in the DNA was visualized by autoradiographic observation of the grains. The DNA strand break appeared as early as 15 min, increased to 10.3-fold at 45 min of 43°C treatment and this level related reciprocally to clonogenicity of the cell. The nick translation method thus provides a rapid in situ assay for determining heat-induced DNA damage of cultured cells, in a semi-quantitative manner.

UR - http://www.scopus.com/inward/record.url?scp=0023934712&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0023934712&partnerID=8YFLogxK

U2 - 10.1016/0304-3835(88)90259-5

DO - 10.1016/0304-3835(88)90259-5

M3 - Article

C2 - 3370628

AN - SCOPUS:0023934712

VL - 40

SP - 33

EP - 38

JO - Cancer Letters

JF - Cancer Letters

SN - 0304-3835

IS - 1

ER -