沉默Ku86基因对Hela细胞放射敏感性影响  

Effects of Ku86 knockdown on the radiosensitivity of Hela cells

作　　者：何科[1] 瞿虎[2] 徐丽南[2] 牛刚[1] 费慧[1] 姜红叶[1] 梁炎春[1] 李婷婷[2] 尧冰 

机构地区：[1]中山大学附属第一医院妇产科,广州510080 [2]中山大学附属第六医院生殖中心,广州510655

出　　处：《中华放射肿瘤学杂志》2016年第9期1008-1012,共5页Chinese Journal of Radiation Oncology

摘　　要：目的探讨siRNA沉默Ku86基因表达对宫颈癌Hela细胞放射敏感性的影响。方法x射线照射后采用qRT—PCR、蛋白印迹法检测Ku86基因在Hela细胞中的表达水平，利用siRNA沉默Ku86基因表达。将si．Nc、si—KU86转染到宫颈癌Hela细胞中，0、2、4、6、8、10GyX射线照射。CCK-8试剂盒检测细胞增殖活性，流式细胞仪检测细胞凋亡情况，克隆形成实验分析转染前后细胞放射敏感性变化，检测p53、Caspase．8表达分析沉默Ku86基因表达对放射诱导的细胞凋亡影响。结果X线照射后Ku86mRNA和蛋白表达水平上调．沉默Ku86表达降低了Hela细胞的增殖能力和克隆形成能力，放射增敏比为1．57。沉默Ku86表达上调了p53和Caspase-8表达，细胞凋亡增加。结论siRNA沉默Ku86表达提高了宫颈癌Hela细胞的放射敏感性。Objective To study the effects of siRNA-mediated Ku86 knockdown on the radiosensitivity of human cervical carcinoma Hela cells. Methods Quantitative real-time PCR and Western blot were used to determine the expression of Ku86 gene in Hela cells exposed to X-ray radiation, siRNA was used for Ku86 knockdown. After transfection with si-NC or si-KU86, Hela cells were exposed to 0, 2, 4, 6, 8, and 10 Gy of X-ray. The CCK-8 kit, flow cytometry, and colony formation assay were used to evaluate cell proliferation, apoptosis, and changes in radiosensitivity after transfection, respectively. The expression of p53 and caspase-8 was evaluated to analyze the effect of Ku86 knockdown on radiation-induced apoptosis. Results After X-ray irradiation, both mRNA and protein expression of Ku86 was upregulated. Ku86 knockdown reduced cell proliferation and colony formation ability. The radiation sensitization enhancement ratio was 1.57. Ku86 knockdown also elevated the expression of p53 and caspase-8 and enhanced apoptosis. Conclusions siRNA-mediated Ku86 knockdown enhances the radiosensitivity of human cervical carcinoma Hela cells.

关 键 词：RNA干扰 Ku86基因 宫颈癌细胞系 放射敏感性 

分 类 号：R737.33[医药卫生—肿瘤] R730.55[医药卫生—临床医学]