Ma Zhihui, Ma Jinyang, Lang Bojuan, Xu Fei, Zhang Bo, Wang Xiangyu
Department of Neurosurgery, The First Hospital of Qinhuangdao, The First Hospital of Qinhuangdao, Qinhuangdao, Hebei, China.
Department of Neurology, The First College of Clinical Medical Sciences, China Three Gorges University & Yichang Central People's Hospital, Yichang, Hubei, China.
Mol Biotechnol. 2023 Mar;65(3):466-476. doi: 10.1007/s12033-022-00540-x. Epub 2022 Aug 22.
This study was aimed at probing into the regulatory effects of circular RNA (circRNA)_0001982 on glioma cell proliferation, migration, invasion, and cell cycle, and its underlying mechanism. CircRNA expression profile of glioma tissues/normal brain tissues was downloaded from the Gene Expression Omnibus (GEO) database and analyzed. Circ_0001982, microRNA (miRNA, miR)-1205, and E2F transcription factor 1 (E2F1) expressions in glioma tissues and cell lines were quantified using quantitative real-time polymerase chain reaction (qRT-PCR) and/or Western blot. Glioma cell proliferation, migration, invasion, and cell cycle were detected employing cell counting kit-8 (CCK-8), 5-Ethynyl-2'-deoxyuridine (EdU), scratch-healing, Transwell, and flow cytometry assays, respectively. The targeting relationships between miR-1205 and circ_0001982, and miR-1205 and E2F1 3'UTR were verified using bioinformatics, dual-luciferase reporter experiments, and RNA immunoprecipitation (RIP) assay. Pearson's correlation analysis was applied to detect the correlations among circ_0001982, miR-1205, and E2F1 expression levels. Circ_0001982 expression level was increased in glioma tissues and correlated with larger tumor size. Circ_0001982 overexpression enhanced glioma cell proliferation, migration, and invasion, and accelerated cell cycle progression while knocking down circ_0001982 exerted opposite effects. Circ_0001982 directly targeted miR-1205, and miR-1205 directly targeted E2F1. Besides, circ_0001982 could up-regulate E2F1 expression via repressing miR-1205 expression. Circ_0001982 accelerates glioma progression by modulating the miR-1205/E2F1 axis.
本研究旨在探讨环状RNA(circRNA)_0001982对胶质瘤细胞增殖、迁移、侵袭及细胞周期的调控作用及其潜在机制。从基因表达综合数据库(GEO)下载并分析胶质瘤组织/正常脑组织的circRNA表达谱。采用定量实时聚合酶链反应(qRT-PCR)和/或蛋白质免疫印迹法检测胶质瘤组织和细胞系中circ_0001982、微小RNA(miRNA,miR)-1205和E2F转录因子1(E2F1)的表达。分别采用细胞计数试剂盒-8(CCK-8)、5-乙炔基-2'-脱氧尿苷(EdU)、划痕愈合、Transwell和流式细胞术检测胶质瘤细胞的增殖、迁移、侵袭及细胞周期。通过生物信息学、双荧光素酶报告基因实验和RNA免疫沉淀(RIP)实验验证miR-1205与circ_0001982以及miR-1205与E2F1 3'非翻译区(UTR)之间的靶向关系。采用Pearson相关分析检测circ_0001982、miR-1205和E2F1表达水平之间的相关性。circ_0001982在胶质瘤组织中的表达水平升高,且与肿瘤体积较大相关。circ_0001982过表达增强了胶质瘤细胞的增殖、迁移和侵袭能力,并加速了细胞周期进程,而敲低circ_0001982则产生相反的效果。circ_0001982直接靶向miR-1205,miR-1205直接靶向E2F1。此外,circ_0001982可通过抑制miR-1205的表达上调E2F1的表达。circ_0001982通过调节miR-1205/E2F1轴促进胶质瘤进展。