A novel parameter, cell-cycle progression index, for radiation dose absorbed estimation in the premature chromosome condensation assay
A novel parameter, cell-cycle progression index, for radiation dose absorbed estimation in the premature chromosome condensation assay
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DOI：
10.1093/rpd/ncu126
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发表时间：
2014-06-01
影响因子：
1
通讯作者：
Blakely, William F.
中科院分区：
文献类型：
--
作者：
Miura, Tomisato;Nakata, Akifumi;Blakely, William F.
由于calyculin A在细胞周期的各个阶段诱导染色体凝聚，因此calyculin A诱导的染色体过早凝聚（PCC）试验是评估细胞中细胞周期分布的一种简单而有用的方法。在这项研究中，一个新的参数，细胞周期进展指数（CPI），在PCC测定被验证为一种新的生物标记物的生物剂量。在获得知情同意后，从健康供体抽取外周血。采用人外周血淋巴细胞（PBL）离体照射（Co-60-γ射线：类似于0.6戈伊min（-1）或X射线：1.0戈伊min（-1）; 0-10戈伊）模型研究CPI。进行Calyculin A诱导的PCC测定用于染色体制备。PCC细胞按细胞周期分期可分为非PCC、G1-PCC、S-PCC、G2/M-PCC和M/A-PCC五类。CPI计算为G2/M-PCC细胞与G1-PCC细胞的比率。PCC阶段分布随照射剂量的变化而显著。G1-PCC细胞分数显着减少，和G2/M-PCC细胞分数增加，在10-Gy照射PBL培养48小时后。CPI水平与γ射线照射[y = 0.6729 + 0.3934 exp（0.5685D），r = 1.0000，p < 0.0001]和X射线照射[y = -0.3743 +0.9744 exp（0.3321D），r = 0.9999，p < 0.0001]拟合成指数剂量-反应曲线。在人外周血离体照射模型中，对CPI没有显著的个体（p = 0.853）或性别效应（p = 0.951）。此外，CPI测量是快速的（每个病例< 15分钟）。这些结果表明，消费物价指数是一个有用的筛选工具，在辐射事件后，特别是在大规模伤亡的辐射事件后的早期辐射照射范围从0到10戈伊的辐射剂量的评估。
The calyculin A-induced premature chromosome condensation (PCC) assay is a simple and useful method for assessing the cell-cycle distribution in cells, since calyculin A induces chromosome condensation in various phases of the cell cycle. In this study, a novel parameter, the cell-cycle progression index (CPI), in the PCC assay was validated as a novel biomarker for biodosimetry. Peripheral blood was drawn from healthy donors after informed consent was obtained. CPI was investigated using a human peripheral blood lymphocyte (PBL) ex vivo irradiation (Co-60-gamma rays: similar to 0.6 Gy min(-1), or X ray: 1.0 Gy min(-1); 0-10 Gy) model. The calyculin A-induced PCC assay was performed for chromosome preparation. PCC cells were divided into the following five categories according to cell-cycle stage: non-PCC, G1-PCC, S-PCC, G2/M-PCC and M/A-PCC cells. CPI was calculated as the ratio of G2/M-PCC cells to G1-PCC cells. The PCC-stage distribution varied markedly with irradiation doses. The G1-PCC cell fraction was significantly reduced, and the G2/M-PCC cell fraction increased, in 10-Gy-irradiated PBL after 48 h of culture. CPI levels were fitted to an exponential dose-response curve with gamma-ray irradiation [y = 0.6729 + 0.3934 exp(0.5685D), r = 1.0000, p < 0.0001] and X-ray irradiation [y = -0.3743 + 0.9744 exp(0.3321D), r = 0.9999, p < 0.0001]. There were no significant individual (p = 0.853) or gender effects (p = 0.951) on the CPI in the human peripheral blood ex vivo irradiation model. Furthermore, CPI measurements are rapid (< 15 min per case). These results suggest that the CPI is a useful screening tool for the assessment of radiation doses received ranging from 0 to 10 Gy in radiation exposure early after a radiation event, especially after a mass-casualty radiological incident.