RNA干扰沉默IGFIR表达对A549细胞放射增敏作用的研究  

Effects of IGF1R RNA interference mediated by lentivirus on radiosensitivity of human lung adenocarcinoma A549 cells

作　　者：王海清[1] 贾晓民[1] 杜永亮[1] 赵杰[1] 徐永红[1] 施萍[1] 

机构地区：[1]徐州医学院第二附属医院呼吸科,221006

出　　处：《国际呼吸杂志》2015年第4期277-280,共4页International Journal of Respiration

基　　金：江苏徐州市科技计划项目（XF11C102）

摘　　要：目的构建靶向胰岛素样生长因子1受体（IGFIR）-siRNA重组慢病毒表达载体，观察其对人肺腺癌A549细胞放疗增敏作用并探讨其机制。方法构建IGF1R—siRNA重组慢病毒，感染A549细胞，蛋白免疫印迹法检测IGFIR沉默效率；荧光实时定量PCR法及ELISA法分别检测缺氧诱导因子1α（HIF-1α）、血管内皮生长因子（VEGF）和Bad基因及蛋白表达情况。克隆形成实验检测放射增敏作用。结果IGFIR—siRNA重组慢病毒使A549细胞IGF1R蛋白沉默效率达70．53％；HIF-1α、VEGF和Bad基因及蛋白表达量均显著降低；A549细胞对放射治疗的敏感性增加，平均致死剂量由1．20Gy下降至0．94Gy，放射增敏比达1．28。结论沉默IGF1R对人肺腺癌A549细胞具有放疗增敏效应，其机制可能与HIF-1α、VEGF和Bad表达下调相关。Objective To construct lentiviral vector targeting insulin-like growth factor-1 receptor （IGF1R）-siRNA,to observe its effects on radiosensitivity of A549 cells, and to discuss the mechanism. Methods The recombinant lentivirus targeting IGF1R-siRNA was constructed and transfected into A549 cells. The efficiency of silencing IGF1R on proteinic level was detected by Western blot. The mRNA expressions of hypoxia inducible factor-1α （HIF-1α）, vascular endothelial growth factor （VEGF）, and Bad were detected by fluorescent real time quantitative PCR. The levels of HIF-1α, VEGF, and Bad protein were detected by ELISA methods. The radiosensitivity was detected by clone formation experiment. Results The efficiency of silencing IGF1R was 70. 53%. The expressions of HIF-1α,VEGF and Bad in genic and proteinic levels reduced significantly. The radiosensitivity of A549 cells enhanced significantly. The Do values declined from 1.20 Gy to 0.94 Gy, and the sensitizing enhancement ratio was 1.28. Conclusions The silencing of IGF1R can enhance the radiosensitivity of A549 cell line. Down-regulated expressions of HIF-1α, VEGF,and Bad could be part of the mechanism.

关 键 词：胰岛素样生长因子1受体 慢病毒载体 放疗增敏 RNA干扰 A549细胞 

分 类 号：R734.2[医药卫生—肿瘤]