CAJ | 학술논문

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,并观察其对萦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RT-PCR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38.52 μg/L降至16.27 μg/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.
목적 탐토만병독개도적RNA간우기술침묵인폐선암A549세포이도소양생장인자1수체(IGF1R)표체,병관찰기대영삼순화료민감성적영향.방법 구건IGF1R중조만병독,감염A549세포,건립은정저표체IGF1R세포주;응용RT-PCR급단백면역인적법검측IGF1R침묵효솔;CCK8비색법검측IGF1R침묵후A549세포대자삼순민감성적영향.구건라서이식류모형,검측자삼순대저표체IGF1R적A549세포이식류생장적억제작용.결과 성공구건IGF1R중조만병독,병건립료은정저표체IGF1R적A549세포주;자삼순대저표체IGF1R적A549세포적반수억제제량유38.52 μg/L강지16.27 μg/L,민감성제고2.37배;저표체IGF1R적A549세포이식류생장완만,여자삼순연합응용후,이식류생장현저억제,체적억류솔체87.5％,중량억류솔체88.2％.결론 만병독개도적IGF1R기인침묵능억제인폐선암세포증식,병현저제고선암세포대자삼순적민감성.
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 IGF1R was conscructed and transfected into A549 cells which expressed IGF1R 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 IGF1R 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.