Determination of 8-oxoguanine in individual cell nucleus of gamma-irradiated mammalian cells.
8-Oxoguanine, through its ability to mispair bases other than cytosine, is assumed to be one of the most potent premutagenic lesions in nuclear DNA damaged by reactive oxygen radicals. In this study, we examine whether the presence of residual 8-oxoguanine can be detected in mammalian cells after exposure to ionizing radiation. MOLT-4 human leukemia cells and CHO-K1 Chinese hamster cells were acutely irradiated in vitro with 0, 0.2, 0.4, 0.6 and 1.0 Gy gamma radiation at room temperature. The amounts of 8-oxoguanine and total DNA in the cell nucleus were detected by fluorescein-isothiocyanate (FITC)-labeled avidin, which binds specifically and directly to 8-oxoguanine, and propidium iodide, respectively. The intensity ratios between these two fluorescent dyes were then taken as indices to measure the content of 8-oxoguanine within individual cells. We found an apparent dose-dependent increase in the amount of 8-oxoguanine accumulated in cells of both lines. Moreover, the content of 8-oxoguanine decreased from 2 to 20 h after irradiation in CHO-K1 cells, which may reflect the time-dependent repair processes at the 8-oxoguanine lesions. This novel approach may provide a sensitive tool for in situ measurement of 8-oxoguanine in cells or even in the human body after exposure to ionizing radiation.
Chen
S K
SK
Institute of Environmental Health Sciences, and Institute of Public Health, National Yangming University, Taipei, Taiwan.
Tsai
M H
MH
Hwang
J J
JJ
Chang
W P
WP
Chan
W P
WP
D000818:Animals; D016466:CHO Cells; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism / Q000528:radiation effects*; D006224:Cricetinae; D004249:DNA Damage; D004260:DNA Repair; D005453:Fluorescence; D006147:Guanine / Q000031:analogs & derivatives* / Q000378:metabolism*; D006801:Humans
C024829:8-hydroxyguanine; D006147:Guanine