| Fan Huadong,Sun Yuxiang,Chen Shaopen,Xu Shaohai,Wu Lijun.The impact of α-particles irradiation on src kinase activity and autophagy system mediated by ROS[J].Chinese Journal of Radiological Medicine and Protection,2015,35(7):481-484,527 |
| The impact of α-particles irradiation on src kinase activity and autophagy system mediated by ROS |
| Received:February 16, 2015 |
| DOI：10.3760/cma.j.issn.0254-5098.2015.07.001 |
| KeyWords:α-particles Autophagy Reactive oxygen species src kinase |
| FundProject:国家自然科学基金(81273004,31470829) |
| Author Name | Affiliation | E-mail | | Fan Huadong | Key Laboratory of Ion Beam Bioengineering, Anhui Province, Chinese Academy of Science, School of Nuclear Science and Technology, University of Science and Technology of China, Hefei 230027, China | | | Sun Yuxiang | Key Laboratory of Ion Beam Bioengineering, Anhui Province, Chinese Academy of Science, School of Nuclear Science and Technology, University of Science and Technology of China, Hefei 230027, China | | | Chen Shaopen | Key Laboratory of Ion Beam Bioengineering, Anhui Province, Chinese Academy of Science, School of Nuclear Science and Technology, University of Science and Technology of China, Hefei 230027, China | | | Xu Shaohai | Key Laboratory of Ion Beam Bioengineering, Anhui Province, Chinese Academy of Science, School of Nuclear Science and Technology, University of Science and Technology of China, Hefei 230027, China | | | Wu Lijun | Key Laboratory of Ion Beam Bioengineering, Anhui Province, Chinese Academy of Science, School of Nuclear Science and Technology, University of Science and Technology of China, Hefei 230027, China | ljw@ipp.ac.cn |
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| Abstract:: |
| Objective To investigate the modulation role of autophagy in radiation-induced cell death by detecting the response of src kinase activity and autophagy in HEK293 cells irradiated with different dose of α-particle. Methods HEK293 cells were irradiated by contral group (0 cGy) a low dose group (10 cGy) and high dose group (300 cGy) α-particles. Molecular probe 2', 7'-dichlorofluorescin (DCFHDA) was used to detect the cell ROS. The src kinase activity and endogenous protein level of LC3I/II were monitored by Western Blot. Cell death rate of irradiated cells pretreated with autophagy inducer of rapamycin was tested by flow cytometry. Results Compared with control group, the ratio of LC3I/II decreased (t=4.07,P<0.05) and the percentage of cells with GFP-LC3 punctuate dots increased (t=12.29,P<0.05) under 10 cGy irradiation, indicating the induction of autophagy. On the contrary, the ratio of LC3I/II increased (t=2.93,P<0.05) and the GFP-LC3 morphology had no obvious change under 300 cGy irradiation. The cellular ROS level reached to the maximum value at 4 h post-irradiation. Both 10 cGy and 300 cGy irradiation could elevate the ROS level (t=17.93,22.88,P<0.05), whereas the amplitude of elevation of 300 cGy irradiation was higher than that of 10 cGy irradiation (t=15.76, 22.66, 14.22, P<0.05). Compared with control group, the 419th site of tyrosine residue in src kinase manifested hyper-phosphorylation (t=5.66,P<0.05) under 10 cGy irradiation whereas it had hypo-phosphorylation under 300 cGy irradiation (t=4.67,P<0.05). Treatment of cells with DMSO could partly restore the impact on src kinase activity under high or low dose irradiation. Pre-treating the cells with autophagy inducer rapamycin could reduce cell death under 300 cGy irradiations(t=12.14,P<0.05). Conclusions High or low dose of α-particles irradiation could inhibit or activate src kinase and autophagy system, respectively. ROS mediated the response of src kinase activity and autophagy system induced by irradiation. Modulation of autophagy could desensitize cell responses to irradiation. |
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