摘要
目的探讨慢病毒介导的RNA干扰技术沉默人肺腺癌A549细胞胰岛素样生长因子1受体（IGF1R）表达，并观察其对紫杉醇化疗敏感性的影响。方法构建IGF1R重组慢病毒，感染A549细胞，建立稳定低表达IGF1R细胞株；应用RTPCR及蛋白免疫印迹法检测IGF1R沉默效率；CCK8比色法检测IGF1R沉默后A549细胞对紫杉醇敏感性的影响。构建裸鼠移植瘤模型，检测紫杉醇对低表达IGF1R的A549细胞移植瘤生长的抑制作用。结果成功构建IGF1R重组慢病毒，并建立了稳定低表达IGF1R的A549细胞株；紫杉醇对低表达IGF1R的A549细胞的半数抑制剂量由38．52μg／L降至16．27μg／L，敏感性提高2．37倍；低表达IGF1R的A549细胞移植瘤生长缓慢，与紫杉醇联合应用后，移植瘤生长显著抑制，体积抑瘤率达87．5％，重量抑瘤率达88．2％。结论慢病毒介导的IGF1R基因沉默能抑制人肺腺癌细胞增殖，并显著提高腺癌细胞对紫杉醇的敏感性。
Objective To discuss insulin-like growth factor-1 receptor （IGF1R） silencing in A549 cells via RNA interference mediated by lentivirus, and to observe its effect on chemosensitivity to paclitaxel. Methods Recombinant lentivirus targeting IGFIR was conseructed and transfected into A549 cells which expressed IGFIR in a low level. The silencing efficiency of IGF1R gene and protein was detected by real time PCR and Western blot. The change of paclitaxel sensitivity after RNA interference was examined by cell counting kit 8 method. Nude mice xenograft tumors model was used to determine the effect of RNA interference targeting IGF1R gene combined with or without paclitaxel on growth inhibition of tumor in vivo. Results The recombinant lentiviral vector expressing shRNA against IGF1R gene was constructed successfully and the cell lines of A549 which expressed IGFIR in a low level were obtained. After RNA interference,the median inhibition concentration of paclitaxel against A549 cells was reduced from 38.52 μg/L. to 16.27 μg/L,and the sensitivity to paclitaxel was increased by 2.37 times. In vivo xenograft tumor model assay showed a significant growth inhibiting effect by RNA interference against IGF1R. When combined with paclitaxel, xenograft tumors were significantly inhibited, and the inhibition rate in volume was 87.5 %, and the inhibition rate in weight was 88.2%. Conclusions RNA interference targeting IGF1R gene could inhibit the proliferation capacity of A549 cell line,and enhance the sensitivity of A549 cells to paclitaxel.
出处
《国际呼吸杂志》
2015年第6期406-409,共4页
International Journal of Respiration
基金
江苏徐州市科技计划项目（XF11C102）
关键词
胰岛素样生长因子1受体
紫杉醇
肺腺癌
慢病毒
Insulin-like growth factor-1 receptor
Paclitaxel
Lung adenocarcinoma
Lentivirus
