Department of Respiratory Medicine, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
Department of Emergency, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
Biomed Pharmacother. 2020 Jul;127:110214. doi: 10.1016/j.biopha.2020.110214. Epub 2020 May 11.
Non-small cell lung cancer (NSCLC) is a type of malignant tumor which threatens human health and life. Recently, some researches on long non-coding RNAs (lncRNAs) in NSCLC has elucidated critical regulatory roles in cell proliferation, migration, and invasion, the relative clinical significance and mechanisms of action are still unclear. This study focuses on the important role of a novel lncRNA LINC00665 in the development of NSCLC. Long intergenic non-protein coding RNA 665 gene (LINC00665) was found through microarray analysis and was measured by real-time quantitative PCR (RT-qPCR). The interactions between LINC00665 and miR-138-5p as well as the interactions between miR-138-5p and E2F3 (E2F transcription factor 3) were explored by bioinformatics analysis and dual-luciferase assays. CCK-8, transwell and mouse xenograft assays were performed to investigate the effects of LINC00665 and miR-138-5p on NSCLC proliferation and invasion. As a result, LINC00665 expression was upregulated in NSCLC lung tissues and cells. Downregulated LINC00665 could arrest A549 and H1299 cell proliferation and invasion in vitro, and this finding was recapitulated in vivo. LINC00665 directly regulated the expression of miR-138-5p. Additionally, E2F3 was one of the targets of miR-138-5p; E2F3 without 3'UTR could reverse the inhibitory effects of downregulated LINC00665 on proliferation and invasion in A549 and H1299 cells. In conclusion, dysregulation of LINC00665 plays a vital role in NSCLC progression, indicating that its downregulation may confer decreased cell proliferation and invasion via the miR-138-5p/E2F3 signaling pathway.
非小细胞肺癌（NSCLC）是一种威胁人类健康和生命的恶性肿瘤。最近，对 NSCLC 中长链非编码 RNA（lncRNA）的一些研究揭示了其在细胞增殖、迁移和侵袭中的关键调节作用，但相对的临床意义和作用机制尚不清楚。本研究聚焦于新型 lncRNA LINC00665 在 NSCLC 发生发展中的重要作用。通过微阵列分析发现长基因间非蛋白编码 RNA 665 基因（LINC00665），并通过实时定量 PCR（RT-qPCR）进行测量。通过生物信息学分析和双荧光素酶报告基因实验探索 LINC00665 与 miR-138-5p 以及 miR-138-5p 与 E2F3（E2F 转录因子 3）之间的相互作用。通过 CCK-8、Transwell 及小鼠异种移植实验研究 LINC00665 和 miR-138-5p 对 NSCLC 增殖和侵袭的影响。结果表明，LINC00665 在 NSCLC 肺组织和细胞中表达上调。下调 LINC00665 可在体外抑制 A549 和 H1299 细胞的增殖和侵袭，这一发现也在体内得到了验证。LINC00665 直接调控 miR-138-5p 的表达。此外，E2F3 是 miR-138-5p 的靶基因之一；没有 3'UTR 的 E2F3 可逆转下调 LINC00665 对 A549 和 H1299 细胞增殖和侵袭的抑制作用。总之，LINC00665 的失调在 NSCLC 的进展中起着至关重要的作用，表明其下调可能通过 miR-138-5p/E2F3 信号通路降低细胞的增殖和侵袭能力。
