CAJ | 학술논문

目的：构建靶向胰岛素样生长因子1受体(IGF1R)-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 IGF1R 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 IGF1R-siRNA 重组慢病毒使 A549细胞 IGFlR 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1.20 Gy 下降至0.94 Gy,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。
목적：구건파향이도소양생장인자1수체(IGF1R)-siRNA 중조만병독표체재체,관찰기대인폐선암 A549세포방료증민작용병탐토기궤제。방법구건 IGF1R-siRNA 중조만병독,감염 A549세포,단백면역인적법검측 IGF1R 침묵효솔；형광실시정량 PCR 법급 ELISA 법분별검측결양유도인자1α(HIF-1α)、혈관내피생장인자(VEGF)화 Bad 기인급단백표체정황。극륭형성실험검측방사증민작용。결과 IGF1R-siRNA 중조만병독사 A549세 포 IGFlR 단백침묵효솔체70.53%；HIF-1α、VEGF화 Bad 기인급단백표체량균현저강저；A549세포대방사치료적민감성증가,평균치사제량유1.20 Gy 하강지0.94 Gy,방사증민비체1.28。결론침묵 IGF1R 대인폐선암 A549세포구유방료증민효응,기궤제가능여 HIF-1α、VEGF 화 Bad 표체하조상관。
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.