钙结合蛋白S100A8在γ射线损伤小鼠巨噬细胞RAW264.7中的作用  

Effect of calcium-binding protein S100A8 in macrophage cell line RAW264.7 of mouse injured by irradiation

作　　者：丛悦[1] 饶亚岚[1] 陈肖华[1] 董波[1] 李峰生[1] 王治东[1] 罗庆良[1] 

机构地区：[1]军事医学科学院放射与辐射医学研究所,北京100850

出　　处：《辐射研究与辐射工艺学报》2010年第5期267-270,共4页Journal of Radiation Research and Radiation Processing

基　　金：国家自然科学基金(30400119)资助

摘　　要：为探索钙结合蛋白S100A8在γ射线对造血免疫系统损伤中的作用,构建S100A8真核表达载体,转染小鼠巨噬细胞RAW264.7,研究S100A8在γ射线诱导RAW264.7细胞周期紊乱和胞内活性氧改变中的作用。运用RT-PCR和细胞荧光免疫方法鉴定增强表达S100A8的RAW264.7稳定细胞克隆。流式技术检测10Gyγ射线照射6h后细胞周期及H2O2刺激后胞内活性氧的变化。结果显示,与转染空质粒的对照细胞相比,增强表达S100A8细胞克隆G0/G1期细胞比例增高,S期细胞减少,不发生G2/M阻滞;并且H2O2作用后细胞中活性氧水平低于转染空白质粒的细胞。因此,增强表达S100A8的RAW264.7细胞与转染空白质粒细胞相比对辐射更具抗性,其可能原因是与增强表达的S100A8分子具有抗氧化功能有关。In order to establish the stable clones of RAW264.7 cells with S100A8 gene overexpression, and directly investigate the role of S100A8 in γ-rays induced hematoiesis/immune system injury, RT-PCR method and im-muno-fluorescence staining analysis were performed to measure the expression of S100A8 mRNA and protein.The cell cycle was detected by flow cytometry at 6 h after irradiation with 10 Gy γ-rays.The level of reactive oxygen intermediates （ROIs） was detected by flow cytometry at 6h after H2O2 treatment.The cell cycle analysis showed that the percentages of RAW264.7/S100A8 cells had an increase in G0/G1 phase and a decrease in S phase, and G2/M arrest didn＇t occur in experimental group while the level of ROIs was lower than that of the negative control cells.These results indicate that radiation resistant of RAW264.7/S100A8 cells may be related to the antioxidant role of over-expressed S100A8 in comparison to RAW264.7/pcDNA3.1＋ cells.

关 键 词：S100A8 Γ射线 细胞周期 活性氧 

分 类 号：R349.64[医药卫生—基础医学] Q691.5[生物学—生物物理学]