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J Korean Soc Ther Radiol Oncol > Volume 21(1); 2003 > Article
The Journal of the Korean Society for Therapeutic Radiology and Oncology 2003;21(1): 75-81.
Radiation-Induced Apoptosis of Lymphocytes in Peripheral Blood
Yoon Kyeong Oh, Tae Bum Lee, Taek Keun Nam, Keun Hong Kee, Cheol Hee Choi
1Department of Therapeutic Radiology, Chosun University Medical School, Gwangju, Korea. ykoh@chosun.ac.kr
2Medical Research Institute, Chosun University Medical School, Gwangju, Korea.
3Department of Pathology, Chosun University Medical School, Gwangju, Korea.
4Department of Pharmacology, Chosun University Medical School, Gwangju, Korea.
This study quantitatively evaluated the apoptosis in human peripheral blood lymphocytes using flow cytometry, and investigated the possibility of using this method, with a small amount of blood, and the time and dose dependence of radiation-induced apoptosis.
Peripheral blood lymphocytes were isolated from the heparinized venous blood of 11 healthy volunteers, 8 men and 3 women, with each 10 ml of blood being divided into 15 samples. The blood lymphocytes were irradiated using a linear accelerator at a dose rate of 2.4 Gy/min, to deliver doses of 0.5, 1, 2 and 5 Gy. The control samples, and irradiated cells, were maintained in culture medium for 24, 48 and 72 hours following the irradiation. The number of apoptotic cells after the in vitro X-irradiation was measured by flow cytometry after incubation periods of 24, 48 and 72 hours. We also observed the apoptotic cells using a DNA fragmentation assay and electron microscopy.
The rate of spontaneous apoptosis increased in relation to the time interval following irradiation (1.761+/-0.161, 3.563+/-0.564, 11.098+/-2.849, at 24, 48, and 72 hours). The apoptotic cells also increased in the samples irradiated with 0.5, 1, 2 and 5 Gy, in a radiation dose and time interval after irradiation manner, with the apoptosis being too great at 72 hours after irradiation. The dose-response curves were characterized by an initial steep increase in the number of apoptotic cells for irradiation doses below 2 Gy, with a flattening of the curves as the dose approached towards 5 Gy.
The flow cytometric assay technique yielded adequate data, and required less than 1 mL of blood. The time and dose dependence of the radiation-induced apoptosis, was also shown. It is suggested that the adequate time interval required for the evaluation of apoptosis would be 24 to 48 hours after blood sampling.
Key Words: Lymphocyte, Blood, Radiation, Apoptosis
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