马季,叶才勇,丁楠,朱佳赟,胡文涛.DNA-PKcs在细胞辐射超敏感性中的作用研究[J].中华放射医学与防护杂志,2018,38(7):489-493
DNA-PKcs在细胞辐射超敏感性中的作用研究
DNA-PKcs functions in cellular hyper-radiosensitivity
投稿时间:2018-03-13  
DOI:10.3760/cma.j.issn.0254-5098.2018.07.002
中文关键词:  肿瘤细胞  低剂量辐射超敏感性  细胞周期检测点激酶2  DNA依赖蛋白激酶催化亚基
英文关键词:Tumor cells  Low dose hyper-radiosensitivity  Chk2 kinase  DNA-PKcs
基金项目:国家自然科学基金(11405235)
作者单位E-mail
马季 215123 苏州大学医学部放射医学与防护学院  
叶才勇 730000 兰州, 中国科学院近代物理研究所空间辐射生物研究室  
丁楠 730000 兰州, 中国科学院近代物理研究所空间辐射生物研究室  
朱佳赟 730000 兰州, 中国科学院近代物理研究所空间辐射生物研究室  
胡文涛 215123 苏州大学医学部放射医学与防护学院 wthu@suda.edu.cn 
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中文摘要:
      目的 研究DNA-PKcs在细胞辐射超敏感性中的作用。方法 X射线照射M059K和M059J细胞后,克隆形成法实验检测其存活分数;微核分析法和γ-H2AX焦点形成实验检测DNA损伤;Western blot实验检测M059K,M059J细胞中磷酸化Chk1、Chk2和总Chk1、Chk2蛋白的表达。结果 在X射线照射剂量<1 Gy时,M059K细胞呈现出辐射超敏感性;DNA损伤水平不能用于表征低剂量区的HRS/IRR;0.2 Gy X射线照射后,M059K细胞中pChk1/Chk1在20~60 min内显著高于M059J细胞(t=14.157、13.661、14.177、11.317、14.512, P<0.05);0.2 Gy X射线照射后,M059K细胞中pChk2/Chk2在20~50 min内显著高于M059J细胞(t=13.182、13.868、14.155、14.477, P<0.05)。结论 DNA-PKcs野生型的人胶质瘤M059K细胞中存在着低剂量辐射超敏感性现象,其发生可能与DNA-PKcs介导的G2/M期检验点相关蛋白激活有关。
英文摘要:
      Objective To explore the functions of DNA-PKcs in cellular low dose hyper-radiosensitivity. Methods Colony-formation assay was used to detect the survival fractions of M059K and M059J cell lines treated by X-ray irradiation. Micronucleus assay and γ-H2AX foci assay were used to measure the radiation-induced DNA damage. Western blot was used to detect the relative expression levels of phospho-Chk1, total Chk1, phospho-Chk2 and total Chk2 of M059K and M059J cells. Results The hyper-radiosensitivity was observed in M059K cells irradiated with X-ray of doses lower than 1 Gy. DNA damage levels did not show HRS/IRR in the cell lines we used. pChk1/Chk1 in M059K cells was significantly increased during 20 min to 60 min after 0.2 Gy X-ray irradiation (t=14.157, 13.661, 14.177, 11.317, 14.512, P<0.05); pChk2/Chk2 in M059K cells was markedly increased during 20 min to 50 min after 0.2 Gy X-ray irradiation (t=13.182, 13.868, 14.155, 14.477, P<0.05). Conclusions M059K cells show the phenomenon of low dose hyper-radiosensitivity, which may be related to activation of proteins in G2/M phase checkpoints regulated by DNA-PKcs.
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