| Zhao Hua,Cai Tianjing,Lu Xue,Tian Mei,Liu Qingjie.The optimized cytokinesis-block assay for radiation-induced nucleoplasmic bridge[J].Chinese Journal of Radiological Medicine and Protection,2021,41(3):178-182 |
| The optimized cytokinesis-block assay for radiation-induced nucleoplasmic bridge |
| Received:July 29, 2020 |
| DOI：10.3760/cma.j.issn.0254-5098.2021.03.004 |
| KeyWords:Nucleoplasmic bridge Cytokinesis-block Cell culture time Cytochalasin-B Micronucleus |
| FundProject:国家自然科学基金（81573081） |
| Author Name | Affiliation | E-mail | | Zhao Hua | Key Laboratory of Radiological Protection and Nuclear Emergency, China CDC, National Institute for Radiological Protection, Chinese Center for Disease Control and Prevention, Beijing 100088, China | | | Cai Tianjing | Key Laboratory of Radiological Protection and Nuclear Emergency, China CDC, National Institute for Radiological Protection, Chinese Center for Disease Control and Prevention, Beijing 100088, China | | | Lu Xue | Key Laboratory of Radiological Protection and Nuclear Emergency, China CDC, National Institute for Radiological Protection, Chinese Center for Disease Control and Prevention, Beijing 100088, China | | | Tian Mei | Key Laboratory of Radiological Protection and Nuclear Emergency, China CDC, National Institute for Radiological Protection, Chinese Center for Disease Control and Prevention, Beijing 100088, China | | | Liu Qingjie | Key Laboratory of Radiological Protection and Nuclear Emergency, China CDC, National Institute for Radiological Protection, Chinese Center for Disease Control and Prevention, Beijing 100088, China | liuqingjie@nirp.chinacdc.cn |
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| Abstract:: |
| Objective To explore the feasibility of the optimized cytokinesis-block (CB) assay on radiation-induced nucleoplasmic bridge (NPB), and to provide a scientific basis for the application of NPB in biological dose estimation. Methods Human peripheral blood in vitro was irradiated with 2 Gy 60Co γ-rays at a dose rate of 1 Gy/min (0 Gy control group). According to the culture time after irradiation, blood samples were divided into group 48, 56, 68 and 72 h. Cytochalasin-B (Cyt-B) with a concentration of 6 μg/ml was added into the samples at 28 h and harvested at 48, 56, 68 and 72 h after irradiation, respectively. On the other hand, the blood samples were treated with different concentration of Cyt-B i.e., 0.6, 1, 2, 6 and 10 μg/ml at the beginning of culture (0 h) and harvested at 68 h after irradiation. The proportion of mononucleated, binucleated and multinucleated cells, radiation-induced NPB and micronucleus (MN) frequencies were analyzed. Results The nuclear division index (NDI) and proportion of binucleated cells at 2 Gy and 0 Gy had tendency of increasing with cell culture time. NPB frequencies (0.023 0-0.033 0/cell) and MN frequencies had no significantly difference (P>0.05). With the increase of Cyt-B concentration, NDI and the proportion of binucleated cells in group 2 Gy and 0 Gy also increased, but NPB frequencies (0.023 0-0.047 0/cell) had no significant difference (P>0.05). MN frequencies of group 10 μg/ml were significantly lower than that of group 6 μg/ml (U=2.74, P<0.01). Conclusions Cell culture time and Cyt-B concentration had no significant influence on radiation-induced NPB frequencies, suggesting that NPB could be obtained by appropriately reducing cell culture time and Cyt-B could be added into blood samples at the beginning of culture. But this protocol reduced the number of cells for further analysis, and thus its feasibility for dose estimation still need to be studied. |
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