抄録
Human leukemic MOLT-4 cells undergo apoptosis after X-irradiation through p53-dependent pathway. In MOLT-4 cells, p53 (wild type) is stabilized and increased after X-irradiation. We have studied the detection and the measurement of p53 accumulation per individual cell by flow cytometry. Irradiated cells were fixed, permeabilized, and analyzed on FACSCalibur (Becton Dickinson). Cells were stained with anti-human p53-PE (clone DO-7). The quantification of immunofluorescence was performed using Quantum Simply Cellular (Bangs Laboratories). Irradiated MOLT-4 cells were monitored for p53 at 1-6 h after irradiation by flow cytometory. The molecules per cell increased after 1h of postirradiation incubation after exposure to 9 Gy, and reached to 3.5 times after 5 h relative to control. Strong correlation was observed between the time course of the cell death induction and the increase in p53 per cell when p53 was quantified in terms of number of molecules per cell and the number was chosen at the peak position of the p53 profile. The method is useful for further study in a cell during the course of developing apoptosis. [J Radiat Res 44:450 (2003)]
