Xu Guodong, Shao Guofeng, Pan Qiaoling, Sun Lebo, Zheng Dawei, Li Minghui, Li Ni, Shi Huoshun, Ni Yiming
Department of Cardiothoracic Surgery, The First Affiliated Hospital, School of Medicine, Zhejiang UniversityHangzhou 310006, Zhejiang, China; Department of Cardiothoracic Surgery, The Affiliated Hospital, Ningbo Medical Center Lihuili Hospital, Ningbo UniversityNingbo 315041, Zhejiang, China.
Department of Cardiothoracic Surgery, The Affiliated Hospital, Ningbo Medical Center Lihuili Hospital, Ningbo University Ningbo 315041, Zhejiang, China.
Am J Transl Res. 2017 Feb 15;9(2):478-488. eCollection 2017.
MicroRNAs (miRNAs) play a critical role in cancer development and progression. Bioinformatics analyses has identified eukaryotic translation initiation factor 5A2 (eIF5A2) as a target of miR-9. In this study, we attempted to determine whether miR-9 regulates non-small cell lung cancer (NSCLC) cell invasion and migration by targeting eIF5A2 We examined eIF5A2 expression using reverse transcription-quantitative PCR (RT-qPCR) and subsequently transfected A549 and NCI-H1299 NSCLC cells with a miR-9 mimic or miR-9 inhibitor to determine the migration and invasive capability of the cells via wound healing assay and Transwell invasion assay, respectively. E-cadherin and vimentin expression was detected with western blotting. The miR-9 mimic significantly reduced NSCLC cell invasive and metastatic ability, and the miR-9 inhibitor enhanced NSCLC cell migration activity, increasing the number of migrated cells. There was no significant difference between the negative control siRNA and miR-9 mimic groups after knockdown of eIF5A2; western blotting showed that miR-9 regulated E-cadherin and vimentin expression. These data show that miR-9 regulates NSCLC cell invasion and migration through regulating eIF5A2 expression. Taken together, our findings suggest that the mechanism of miR-9-regulated NSCLC cell invasion and migration may be related to epithelial-mesenchymal transition.
微小RNA(miRNA)在癌症的发生和发展中起着关键作用。生物信息学分析已确定真核翻译起始因子5A2(eIF5A2)是miR-9的一个靶标。在本研究中,我们试图确定miR-9是否通过靶向eIF5A2来调节非小细胞肺癌(NSCLC)细胞的侵袭和迁移。我们使用逆转录定量PCR(RT-qPCR)检测eIF5A2的表达,随后分别用miR-9模拟物或miR-9抑制剂转染A549和NCI-H1299 NSCLC细胞,通过伤口愈合试验和Transwell侵袭试验来确定细胞的迁移和侵袭能力。通过蛋白质印迹法检测E-钙黏蛋白和波形蛋白的表达。miR-9模拟物显著降低了NSCLC细胞的侵袭和转移能力,而miR-9抑制剂增强了NSCLC细胞的迁移活性,增加了迁移细胞的数量。在敲低eIF5A2后,阴性对照小干扰RNA和miR-9模拟物组之间没有显著差异;蛋白质印迹法显示miR-9调节E-钙黏蛋白和波形蛋白的表达。这些数据表明,miR-9通过调节eIF5A2的表达来调节NSCLC细胞的侵袭和迁移。综上所述,我们的研究结果表明,miR-9调节NSCLC细胞侵袭和迁移的机制可能与上皮-间质转化有关。