Cell sorting analysis of cell cycle-dependent X-ray sensitivity in end joining-deficient human cells
Cell sorting analysis of cell cycle-dependent X-ray sensitivity in end joining-deficient human cells
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DOI：
10.1016/j.bbrc.2008.05.147
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发表时间：
2008-08-08
影响因子：
3.1
通讯作者：
Date, Takayasu
中科院分区：
文献类型：
--
作者：
Iwabuchi, Kuniyoshi;Hashimoto, Mitsumasa;Date, Takayasu
非同源末端连接（NHEJ）在电离辐射诱导的DNA双链断裂（DSB）修复中起着重要作用，尤其是在细胞周期的G1期。使用流式细胞术细胞分选仪，我们根据大小分级G1期和S/G2期细胞，以评估NHEJ因子缺陷型DT 40和Nalm-6细胞系中的DSB修复活性。集落形成试验表明，G1富集群体的X射线敏感性正确反映了DT 40和Nalm-6细胞系的DSB修复活性。此外，如通过γ-H2 AX焦点形成所评估的，分选的细胞表现出比化学同步化的细胞更少的DNA损伤。鉴于它不使用荧光标记或化学试剂，这种细胞分选方法比其他方法更简单，毒性更低，使其适用于各种细胞系，包括那些无法通过标准化学处理同步的细胞系。(c)2008年爱思唯尔公司All rights reserved.
Non-homologous end joining (NHEJ) plays a major role in the repair of ionizing radiation-induced DNA double-strand breaks (DSBs), especially during the G1-phase of the cell cycle. Using a flow cytometric cell sorter, we fractionated G1- and S/G2-phase cells based on size to assess the DSB-repair activity in NHEJ factor-deficient DT40 and Nalm-6 cell lines. Colony formation assays revealed that the X-ray sensitivities of the G1-enriched populations correctly reflected the DSB-repair activities of both the DT40 and Nalm-6 cell lines. Furthermore, as assessed by gamma-H2AX foci formation, the sorted cells exhibited less DNA damage than chemically synchronized cells. Given that it does not use fluorescent labeling or chemical agents, this method of cell sorting is simpler and less toxic than other methods, making it applicable to a variety of cell lines, including those that cannot be synchronized by standard chemical treatments. (c) 2008 Elsevier Inc. All rights reserved.