Radiation-induced apoptosis in HL60 cells: oxygen effect, relationship between apoptosis and loss of clonogenicity, and dependence of time to apoptosis on radiation dose.

Apoptosis in HL60 human leukemia cells irradiated in vitro was quantified using a DNA fragmentation assay. Dose-response curves for induction of apoptosis in HL60 cells 6 h after irradiation with 280 kVp X rays in air and hypoxia give an oxygen enhancement ratio (OER) of 2.7. This is similar to the OER of 2.8 obtained from survival curves for HL60 cells using a soft agar clonogenic assay. However, HL60 cells are much more sensitive to radiation-induced loss of clonogenicity than to induction of apoptosis at 6 h. For example, 12 Gy in air reduces the surviving fraction to about 0.002 in a clonogenic assay, but 12 Gy does not cause any significant increase in the percentage of apoptosis-like DNA fragmentation 6 h after irradiation compared to unirradiated controls. However, if apoptosis is assayed 2-4 days after irradiation, the HL60 cells show greater sensitivity, with 5 Gy in air causing 45-50% apoptosis at 3 days. When apoptosis is measured 3 days after irradiation, the OER is similar to that obtained for survival and for apoptosis at 6 h. Although the HL60 cells exhibit radiation-induced apoptosis if one waits 2-4 days after low doses of radiation, rather than just 6 h, to conduct the assay, the amount of cells undergoing apoptosis is still not sufficient to account for all the loss of clonogenicity seen when HL60 cells are exposed to ionizing radiation.