| 何为,周福祥,徐会,等.A549细胞线粒体DNA缺失模型建立及其放射敏感性变化[J].中华放射医学与防护杂志,2014,34(4):255-258.He Wei,Zhou Fuxiang,Xu Hui,et al.Establishment of mitochondrial DNA depleted A549 cell line and the change of its radiosensitivity[J].Chin J Radiol Med Prot,2014,34(4):255-258 |
| A549细胞线粒体DNA缺失模型建立及其放射敏感性变化 |
| Establishment of mitochondrial DNA depleted A549 cell line and the change of its radiosensitivity |
| 投稿时间：2013-07-14 |
| DOI：10.3760/cma.j.issn.0254-5098.2014.04.004 |
| 中文关键词: 线粒体DNA缺失 ρ0细胞 mtDNA A549 放射敏感性 |
| 英文关键词:Mitochondrial DNA deletion ρ0 cell mtDNA A549 Radiosensitivity |
| 基金项目:国家自然科学基金（81172129） |
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| 中文摘要: |
| 目的 建立肺癌A549细胞线粒体DNA（mtDNA）缺失模型（ρ0细胞），观察其放射敏感性变化。 方法 用含微量溴化乙锭（EB）的特殊培养液持续培养正常的A549细胞（ρ+细胞）以获得mtDNA完全缺失的ρ0细胞，利用生长缺陷及PCR鉴定该细胞模型。利用克隆形成实验分别检测两种细胞的放射敏感性，利用碘化丙啶（PI）染色法及二氯荧光素双醋酸盐（DCFH-DA）染色法，分析两种细胞在6 MV X射线照射后的细胞周期及活性氧簇（ROS）变化情况。 结果 成功建立A549 ρ0细胞。ρ0细胞较ρ+细胞放射敏感性降低（t=12.57，P<0.01）。放射照射8 Gy后，ρ0细胞较ρ+细胞G2期阻滞时间延长，G2期细胞比例降低（t=6.82，P<0.01），ρ0细胞ROS升高水平明显低于ρ+细胞（t=14.51，P<0.01）。 结论 ρ0细胞较ρ+细胞放射敏感性降低，其机制可能与放射照射后ROS产量降低及G2期阻滞延长有关。 |
| 英文摘要: |
| Objective To establish the mitochondrial DNA depleted cell line (ρ0 cells) of lung cancer cell A549 and to observe the radiosensitivity difference between ρ0 cells and normal A549 cells (ρ+ cells). Methods The ρ0 cells were depleted of mitochondrial DNA by culturing chronically in the presence of low concentration of ethidium bromide (EB), and then the cell model was confined.Radiosensitivity of both ρ0 cells and ρ+ cells was detected using the cologenic formation assay. After 6 MV X-ray irradiation in vitro, cell cycle distribution and reactive oxygen species (ROS) level were analyzed by flow cytometry and fluorescence microplate reader, respectively. Results A ρ0 cell line was successfully established and had a lower radiosensitivity than ρ+ cells (t=12.57, P<0.01). After irradiation with a dose of 8 Gy, compared to ρ+ cells, ρ0 cells showed prolonged G2 arrest with less cells in G2 (t=6.82, P<0.01) and had lower increase of ROS level (t=14.51, P<0.01). Conclusions ρ0 cells have a lower radiosensitivity than ρ+ cells, in which the reduction of ROS production and the prolongation of G2 arrest post-irradiation may be involved. |
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