Radiation sensitivity and the status of some radiation sensitivity markers in relatively sensitive lymphoid cells

The most commonly used indicators of ionizing radiation exposure are cytogenetic measures and survival parameters. All these methods have their advantages, disadvantages and uncertainties, such that better biological estimators of the absorbed dose, especially in the low dose range, are being sought. In this study we analyzed apoptosis and several proteins involved in the regulation of apoptosis as possible indicators of irradiation after relatively small doses (0.1-2 Gy) of X-rays. The studies were carried out in seven lymphoid cell lines: two mouse lymphoma L5178Y, the human pre-B cell leukemia Reh, and four human Epstein-Barr virus-transformed lymphoid cell lines (two apparently normal and two Ataxia-telangiectasia (AT)). We detected apoptosis with the in situ terminal deoxynucleotidyl transferase assay and flow cytometry, and measured the expression of several apoptotic-regulatory proteins (Bcl-2, Bax, Bclx, NF kappa B) with Western blotting. The cytokinesis-block micronucleus assay, comet assay as a measure of DNA damage, and trypan blue survival test were also done for comparison Although for the most of examined parameters of radiation sensitivity: i.e. micronucleus assay, trypan blue test and percentage of apoptosis--there were observed clear dose-effect relationships for all cell lines examined, we did not find agreement between values for these measured parameters. There are marked differences in both timing of apoptosis and percentage of apoptotic cells. Variation in the apoptotic fraction in the controls for different sets of experiments is not very pronounced. There is however considerable variation for the same parameters in irradiated cells, possibly due to their cell cycle status during irradiation, as the cultures were not synchronized. Overall, neither the numbers of apoptotic cells nor the expression of apoptosis-related proteins, nor DNA repair can serve as dose estimators or sensors for these lines, but still these parameters can give valuable supplementary information about radiation sensitivity.