| ZHUANG Liang,YU Shi-ying,HUANG Xiao-yuan.Construction of cell model of silenced Ku80 and the radiobiology change of HeLa cell[J].Chinese Journal of Radiological Medicine and Protection,2007,27(5):447-450 |
| Construction of cell model of silenced Ku80 and the radiobiology change of HeLa cell |
| Received:September 04, 2006 |
| DOI： |
| KeyWords:Ku80 RNA interference Stable transfection Radiosensitivity Cell cycle |
| FundProject: |
| Author Name | Affiliation | E-mail | | ZHUANG Liang | Cancer Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China | | | YU Shi-ying | Cancer Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China | syyu@tjh.tjmu.edu.cn | | HUANG Xiao-yuan | Cancer Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China | | | 熊慧华 | Cancer Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China | | | 熊华 | Cancer Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China | | | 李小兰 | Cancer Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China | | | 冷彦 | Cancer Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China | |
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| Abstract:: |
| Objective To construct the cell model of Ku80 with expression inhibited by siRNA and to explore the role of Ku80 in radiobiology. Methods Ku80-siRNA expression plasmids were constructed and HeLa cells were transfected with these plasmids by lipofectamine. Western blotting was used to measure the expression of Ku80. After irradiation with 6 MV X ray,cells were collected and analyzed by flow cytometry for apoptosis and cell cycle at 24, 48 and 72 h; The radiobiology parameters of four cell lines were acquired by clone formation array. Results Three stable transfected cell clones were obtained, and the inhibition rates of Ku80 protein expression of two positive clones were 89.3% and 96.4%; The apoptosis rates of HeLa cells Ku80 inhibited were higher than control cells at 48 and 72 after X ray irradiation (P＜0.05), but the differences of cell cycle change after irradiation were not significant (P＞0.05). HeLa cells of silenced Ku80 had lower SF2 and D0 than control cells, and their SER (sensitization enhancement ratio) based on D10 were 1.315 and 1.365, respectively. Conclusions The HeLa cell models with Ku80 expression suppressed were successfully established; the inhibition of Ku80 by siRNA could enhance the radiosensitivity of HeLa cells. |
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