Li Fuliang, Wen Jingjing, Shi Jinsheng, Wang Yun, Yang Feifei, Liu Chunying
Department of Pathology, Anqiu People's Hospital, Anqiu, Shandong 262100, P.R. China.
Department of Pathology, Weifang Yidu Central Hospital, Weifang, Shandong 262500, P.R. China.
Exp Ther Med. 2019 Aug;18(2):1175-1183. doi: 10.3892/etm.2019.7668. Epub 2019 Jun 13.
The aberrant expression of microRNAs (miRs) may be involved in tumor growth and progression in human non-small cell lung carcinoma (NSCLC). The present study aimed to investigate the potential roles of miR-191 in NSCLC. Western blotting and reverse transcription-quantitative polymerase chain reaction were performed to assess protein and/or mRNA levels. Scratch wound healing and transwell assays were performed to determine the NSCLC cell migration and invasion. A luciferase demonstrated that CCAAT/enhanced binding protein β (C/EBPβ) was a target of miR-191. Previously, miR-191 has been reported to act as an oncogenic player in multiple human cancers. C/EBPβ has been identified as a target gene of miR-191; however, the roles and underlying mechanisms of miR-191 associated with the regulation of tumor invasion in NSCLC remain unknown. In the present study, it was demonstrated that miR-191 expression levels were higher in human NSCLC tumors compared with in normal adjacent tissue and elevated miR-191 expression levels were closely associated with tumor node metastasis stage in patients with NSCLC. Furthermore, transfection with miR-191 mimic inhibited C/EBPβ expression at the mRNA and protein levels and promoted A549 cell migration and invasion. C/EBPβ was reported to be the direct target gene of miR-191 using a dual luciferase reporter assay. Finally, C/EBPβ siRNA can mimic the effects of miR-191. These findings indicated that miR-191 may function as an oncogene in NSCLC, at least partially due to its negative regulatory on C/EBPβ.
微小RNA(miR)的异常表达可能参与人类非小细胞肺癌(NSCLC)的肿瘤生长和进展。本研究旨在探讨miR-191在NSCLC中的潜在作用。采用蛋白质印迹法和逆转录-定量聚合酶链反应来评估蛋白质和/或mRNA水平。进行划痕伤口愈合实验和Transwell实验以确定NSCLC细胞的迁移和侵袭能力。荧光素酶报告实验表明CCAAT增强子结合蛋白β(C/EBPβ)是miR-191的靶标。此前,已有报道称miR-191在多种人类癌症中发挥致癌作用。C/EBPβ已被确定为miR-191的靶基因;然而,miR-191在NSCLC中与肿瘤侵袭调控相关的作用及潜在机制仍不清楚。在本研究中,结果表明与癌旁正常组织相比,人类NSCLC肿瘤中miR-191的表达水平更高,且miR-191表达水平升高与NSCLC患者的肿瘤淋巴结转移分期密切相关。此外,转染miR-191模拟物可在mRNA和蛋白质水平抑制C/EBPβ的表达,并促进A549细胞的迁移和侵袭。使用双荧光素酶报告实验证实C/EBPβ是miR-191的直接靶基因。最后,C/EBPβ siRNA可模拟miR-191的作用。这些发现表明,miR-191可能在NSCLC中发挥癌基因作用,至少部分是由于其对C/EBPβ的负调控作用。