Liu Cheng, Li Qun-Gen, Zhou Yang, Cao Ying-Yue, Wei Zi-Xin, Jin Ying-Hua, Wang Xin, Chen Ying-Ying, Qi Li, Geng Jian-Xiong, Liu Fang
Department of Thoracic Surgery, The Fourth Affiliated Hospital of Harbin Medical University Harbin 150001, Heilongjiang Province, P. R. China.
Department of Cardiothoracic Surgery, Heilongjiang Provincial Hospital Affiliated to Harbin Institute of Technology Harbin 150001, Heilongjiang Province, P. R. China.
Am J Cancer Res. 2021 Oct 15;11(10):4844-4865. eCollection 2021.
Non-small cell lung cancer (NSCLC) is one type of the most common cancers, which results in the major death worldwide. This study focuses on the understanding of the molecular mechanism of lncRNA NR2F2-AS1 and its regulation on epithelial-mesenchymal transition (EMT) in the development of NSCLC. Expressions of lncRNA NR2F2-AS1, miR-545-5p, c-Met, biliverdin reductase (BVR), ATF-2 and EMT-related markers in NSCLC tissues and cells were measured by western blotting and RT-qPCR assays. The impact of lncRNA NR2F2-AS1 and miR-545-5p on the cell proliferation, migration, invasion and EMT were analyzed by CCK-8, colony formation, wound healing and transwell assays. The interactions among lncRNA NR2F2-AS1, miR-545-5p and c-Met predicted by bioinformatic analysis were evaluated through dual luciferase reporter assay and fluorescence in situ hybridization (FISH). After generating tumor xenografts, immunohistochemistry was utilized to measure the expression of Ki-67 and EMT-related proteins . Our results showed that lncRNA NR2F2-AS1, c-Met, BVR and ATF-2 were overexpressed while miR-545-5p was silenced in NSCLC tissues and cells. Silencing of lncRNA NR2F2-AS1 or upregulating miR-545-5p significantly inhibited the cell proliferation, migration, invasion and EMT process. The EMT process could be inhibited by suppressing c-Met/BVR/ATF-2 axis. The tumor xenograft experiments demonstrated that the tumor growth and EMT process were significantly inhibited by silencing lncRNA NR2F2-AS1 or overexpression of miR-545-5p . LncRNA NR2F2-AS1 promoted the NSCLC development through suppressing miR-545-5p to activate EMT process through c-Met/BVR/ATF-2 axis. Our study indicated that lncRNA NR2F2-AS1 and miR-545-5p could be used as potential therapeutic targets to improve NSCLC treatment.
非小细胞肺癌（NSCLC）是最常见的癌症类型之一，是全球主要的致死病因。本研究旨在了解长链非编码RNA NR2F2-AS1的分子机制及其在NSCLC发生发展过程中对上皮-间质转化（EMT）的调控作用。通过蛋白质免疫印迹法和逆转录-定量聚合酶链反应（RT-qPCR）检测NSCLC组织和细胞中lncRNA NR2F2-AS1、miR-545-5p、c-Met、胆红素还原酶（BVR）、活化转录因子2（ATF-2）及EMT相关标志物的表达。采用细胞计数试剂盒-8（CCK-8）、集落形成、伤口愈合和Transwell实验分析lncRNA NR2F2-AS1和miR-545-5p对细胞增殖、迁移、侵袭及EMT的影响。通过双荧光素酶报告基因实验和荧光原位杂交（FISH）评估生物信息学分析预测的lncRNA NR2F2-AS1、miR-545-5p和c-Met之间的相互作用。构建肿瘤异种移植模型后，采用免疫组织化学法检测Ki-67和EMT相关蛋白 的表达。结果显示，lncRNA NR2F2-AS1、c-Met、BVR和ATF-2在NSCLC组织和细胞中高表达，而miR-545-5p表达下调。沉默lncRNA NR2F2-AS1或上调miR-545-5p可显著抑制细胞增殖、迁移、侵袭及EMT过程。抑制c-Met/BVR/ATF-2轴可抑制EMT过程。肿瘤异种移植实验表明，沉默lncRNA NR2F2-AS1或过表达miR-545-5p可显著抑制肿瘤生长和EMT过程。lncRNA NR2F2-AS1通过抑制miR-545-5p，激活c-Met/BVR/ATF-2轴，促进NSCLC的发生发展。本研究表明，lncRNA NR2F2-AS1和miR-545-5p可作为潜在治疗靶点，改善NSCLC的治疗效果。
