Nick translation detection in situ of cellular DNA strand break induced by radiation
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 (/sup 3/H)-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.
- Research Organization:
- Kyushu Univ., Fukuoka (Japan)
- OSTI ID:
- 6267728
- Journal Information:
- Am. J. Pathol.; (United States), Vol. 134:1
- Country of Publication:
- United States
- Language:
- English
Similar Records
T98G glioma cells have nicks in DNA in quiescent phase
Induction and repair of DNA strand breaks and 1-beta-D-arabinofuranosylcytosine-detectable sites in 40-75 kVp X-irradiated compared to /sup 60/Co gamma-irradiated human cell lines
Related Subjects
59 BASIC BIOLOGICAL SCIENCES
DNA
STRAND BREAKS
HELA CELLS
GENETIC RADIATION EFFECTS
RADIOINDUCTION
AUTORADIOGRAPHY
DNA POLYMERASES
DOSE-RESPONSE RELATIONSHIPS
ESCHERICHIA COLI
TRITIUM COMPOUNDS
BACTERIA
BIOLOGICAL EFFECTS
BIOLOGICAL RADIATION EFFECTS
ENZYMES
GENETIC EFFECTS
LABELLED COMPOUNDS
MICROORGANISMS
NUCLEIC ACIDS
NUCLEOTIDYLTRANSFERASES
ORGANIC COMPOUNDS
PHOSPHORUS-GROUP TRANSFERASES
POLYMERASES
RADIATION EFFECTS
TRANSFERASES
560120* - Radiation Effects on Biochemicals
Cells
& Tissue Culture
550401 - Genetics- Tracer Techniques