Department of Oncology, Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan, China.
Department of Digestive Endoscopy, Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan, China.
Thorac Cancer. 2020 Aug;11(8):2224-2236. doi: 10.1111/1759-7714.13529. Epub 2020 Jun 11.
Non-small cell lung cancer (NSCLC) is one of the leading causes of cancer-related death globally. This study aimed to disclose the role of circular RNA circ_0072088 in NSCLC and illustrate its potential mechanism.
Quantitative real-time polymerase chain reaction (qRT-PCR) was applied to detect the expression of circ_0072088, zinc finger RNA binding protein (ZFR), microRNA-377-5p (miR-377-5p) and NOVA alternative splicing regulator 2 (NOVA2). The viability, colony formation, cell cycle, migration and invasion of NSCLC cells were measured by cell counting kit-8 (CCK8) assay, colony formation assay, flow cytometry, wound healing assay and transwell invasion assay. Western blot assay was employed to examine the protein levels of proliferating cell nuclear antigen (PCNA), Cyclin D1, matrix metallopeptidase 2 (MMP2), MMP9 and NOVA2. The downstream targets of circ_0072088 and miR-377-5p were searched through using circular RNA Interactome and TargetScan databases, and the interaction between miR-377-5p and circ_0072088 or NOVA2 was confirmed by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. in vivo tumor growth assay was used to evaluate the functions of circ_0072088 in the progression of NSCLC in vivo.
GSE101586 dataset and the analysis of tissue specimens showed that circ_0072088 was aberrantly upregulated in tumor tissues of lung cancer and NSCLC. Circ_0072088 interference caused marked suppression on the proliferation and motility of NSCLC cells. Circ_0072088 could negatively regulate miR-377-5p through direct combination. Circ_0072088 contributed to the progression of NSCLC through sponging miR-377-5p. MiR-377-5p could directly interact with NOVA2, and the overexpression of NOVA2 overturned miR-377-5p-mediated influence on NSCLC cells. Circ_0072088 facilitated the progression of NSCLC in vivo.
Circ_0072088 facilitated the proliferation and metastasis of NSCLC cells through upregulating NOVA2 via functioning as a competitive endogenous RNA (ceRNA) for miR-377-5p.
非小细胞肺癌（NSCLC）是全球癌症相关死亡的主要原因之一。本研究旨在揭示环状 RNA circ_0072088 在 NSCLC 中的作用，并阐明其潜在机制。
采用实时定量聚合酶链反应（qRT-PCR）检测 circ_0072088、锌指 RNA 结合蛋白（ZFR）、微小 RNA-377-5p（miR-377-5p）和 NOV 剪接调节因子 2（NOVA2）的表达。通过细胞计数试剂盒-8（CCK8）测定、集落形成测定、流式细胞术、划痕愈合测定和 Transwell 侵袭测定检测 NSCLC 细胞的活力、集落形成、细胞周期、迁移和侵袭。采用 Western blot 测定法检测增殖细胞核抗原（PCNA）、细胞周期蛋白 D1、基质金属蛋白酶 2（MMP2）、MMP9 和 NOVA2 的蛋白水平。通过环状 RNA 相互作用组和 TargetScan 数据库搜索 circ_0072088 和 miR-377-5p 的下游靶标，并通过双荧光素酶报告基因测定和 RNA 免疫沉淀（RIP）测定验证 miR-377-5p 与 circ_0072088 或 NOVA2 之间的相互作用。体内肿瘤生长试验用于评估 circ_0072088 在体内 NSCLC 进展中的作用。
GSE101586 数据集和组织标本分析表明，circ_0072088 在肺癌和 NSCLC 肿瘤组织中异常上调。circ_0072088 干扰导致 NSCLC 细胞增殖和迁移明显受到抑制。circ_0072088 可以通过直接结合负调控 miR-377-5p。circ_0072088 通过海绵 miR-377-5p 促进 NSCLC 的进展。miR-377-5p 可以直接与 NOVA2 相互作用，而 NOV2 的过表达推翻了 miR-377-5p 对 NSCLC 细胞的影响。circ_0072088 促进了体内 NSCLC 的进展。
circ_0072088 通过作为 miR-377-5p 的竞争性内源 RNA（ceRNA）上调 NOV2，促进 NSCLC 细胞的增殖和转移。
