RNAi 介导 IGF-1R 基因沉默对非小细胞肺癌PC-9／ZD 细胞株 EMT 的影响

摘要

Objective To investigate the effect of insulin-like growth factor 1 receptor ( IGF-1R) gene silencing medi-ated by RNA interference ( RNAi) on cell epithelial-mesenchymal transition ( EMT) in non-small-cell lung cancer PC-9/ZD cells.Methods The recombinant lentivirus specific to IGF-1R gene was constructed to be transfected into EGFR-TKIs-resistant non-small-cell lung cancer PC-9/ZD cells.Real-time quantitative RT-PCR was used to detect the inhibition rate of IGF-1R in PC-9/ZD cells;meanwhile, after IGF-1R was inhibited, the mRNA expression of E-cadherin and Vimen-tin was detected , and the invasive and migratory capabilities of cells were determined by Transwell assay .Results The RNAi recombinant lentivirus targeting IGF-1R was successfully constructed .The final titer acquired was 1 ×106 TU/μL, and the inhibition rate of IGF-1R mRNA was 71.4%.After the IGF-1R gene was inhibited, the PC-9/ZD cells displayed epithelial phenotypes, and their capabilities of invasion and migration were significantly decreased (P<0.05).The ex-pression of epithelial cell marker E-cadherin mRNA was increased (P<0.05), accompanied by down-regulation of Vimen-tin mRNA level (P<0.05).Conclusion IGF-1R silencing can reverse cell EMT in non-small-cell lung cancer PC-9/ZD cells.%目的：探讨RNA干扰（ RNAi）介导胰岛素样生长因子1受体（ IGF-1R）基因沉默对非小细胞肺癌（NSCLC）细胞PC-9/ZD发生上皮-间质转化（EMT）的影响。方法构建靶向IGF-1R基因的RNAi重组慢病毒，感染NSCLC表皮生长因子受体-酪氨酸激酶抑制剂（ EGFR-TKIs）获得性耐药细胞PC-9/ZD；实时荧光定量PCR法检测IGF-1R表达抑制效应及IGF-1R基因沉默后EMT相关分子的mRNA表达变化，Transwell实验检测细胞侵袭转移能力变化。结果成功构建靶向IGF-1R基因的RNAi重组慢病毒，滴度为1×106 TU/μL，对PC-9/ZD细胞IGF-1R基因的mRNA抑制效率为71．4％。 IGF-1R基因沉默后，PC-9/ZD细胞形态呈现间质向上皮化转变，上皮标志物E-cadherin的mRNA表达量升高（P＜0．05），间质标志物Vimentin的mRNA表达量降低（P＜0．05），且侵袭转移能力减弱（P＜0．05）。结论靶向IGF-1R基因沉默可逆转PC-9/ZD细胞EMT。