利用小干扰RNA沉默长非编码RNA ZFAS1能够促进安罗替尼对非小细胞肺癌的抗肿瘤活性

siRNA of lncRNA ZFAS1 Enhances the Sensitivity of Non-Small Cell Lung Cancer Cells to Anlotinib

目的:利用小干扰RNA(si RNA)下调长非编码RNA(lnc RNAs)ZFAS1,探索靶向ZPAS1的安罗替尼(anlotinib)增敏治疗策略。方法:在非小细胞肺癌细胞A549中转染对照si RNA或ZFAS1的小干扰RNA(si ZFAS1)后,将细胞注射入裸鼠皮下形成裸鼠皮下肿瘤,对动物口服灌胃给予安罗替尼,确定下调ZFAS1的小干扰RNA对安罗替尼抑制A549细胞皮下肿瘤的影响。进一步将A549细胞通过肝门静脉注射进入裸鼠肝脏,确定下调ZFAS1的小干扰RNA对安罗替尼抑制A549细胞在裸鼠肝脏原位形成肿瘤的影响。收集细胞样品,检测si ZFAS1对N-钙黏蛋白、E-钙黏蛋白及波形蛋白等NOTCH下游上皮-间质转化相关指示分子表达的影响。结果:在A549细胞中转染ZFAS-1的小干扰RNA能够抑制A549细胞的上皮间质转化作用,促进高安罗替尼对A549细胞的杀伤作用。结论:ZFAS1是非小细胞肺癌分子靶向治疗的潜在干预靶标,利用小干扰RNA能够促进安罗替尼对A549细胞的杀伤作用。

Objective： To examine whether down-regulation of ZDAS1, a long non-coding RNA（lncRNA） mediating the activation of NOTCH pathway, could enhance the sensitivity of non-small cell lung cancer（NSCLC） cells to anlotinib, a novel molecular targeted agent. Methods： A549 cells were cultured and transfected with control small interfering RNA（siRNA） or ZFAS1 siRNA（siZFAS1）. Then, cells were treated by indicated concentration of anlotinib for MTT analysis. Next, A549 cells were injected into nude mice to form the subcutaneous tumor models or lung cancer liver metastasis model. After A549 seeding, nude mice were treated with anlotinib to examine the effect of ZFAS1 siRNA on anlotinib. The epithelial-mesenchymal transition（EMT） process of A549 cells was detected by examining the expression of EMT indicators, e.g. E-cadherin, N-cadherin or vimentin. Results： Transfec- tion of siZFAS1 but not control siRNA enhanced tile antitumor effect of anlotinib on A549 cell, the IC50 value of anlotinib decreased from 0.50±0.04 to 0.14±0.01 μmol/L. Transfection of siZFAS1 also enhanced the anti-tumor effect of anlotinib on the subcutaneous growth or intrahepatic metastasis of A549 cells. Conclusion： Knockdown of ZFAS1 could enhance the antitumor efficiency of anlotinib on NSCLC cells.