Li Huibing, Jin Xin, Lai Mingyao, Li Yongshi, Li Ruixing, Yang Huihui, Yang Baoying
Department of Neurosurgery, Guangdong 999 Brain Hospital, Guangzhou, Guangdong Province, PR China.
Department of Ultrasonography, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong Province, PR China.
J Neurogenet. 2023 Sep;37(3):93-102. doi: 10.1080/01677063.2023.2199067. Epub 2023 Apr 27.
A great amount of reaches have confirmed that circular RNAs (circRNAs) are novel regulators in glioma progression. Here, our work aimed to probe the specific role of circ_CLIP2 in glioma. The mRNA and protein expressions were analyzed by qRT-PCR and western blot, respectively. Cell viability, migration, invasion and apoptosis were examined by MTT assay, tranwell and flow cytometry assays, respectively. Moreover, the binding relationships between circ_CLIP2, microRNA (miR)-641 and erythropoietin-producing human hepatocellular (Eph)A3 were verified by dual luciferase reporter gene assay and/or RIP assay. The following data showed that circ_CLIP2 and EPHA3 were markedly increased in glioma tissues and cells, while miR-647 was downregulated. Gain- and loss-of-function experiments discovered that circ_CLIP2 knockdown remarkably inhibited cell proliferation, migration and invasion and promoted cell apoptosis of glioma cells, while these effects of circ_CLIP2 knockdown were abolished by miR-641 inhibition. Circ_CLIP2 was proved as a sponge of miR-641 to competitively upregulate EPHA3 expression. In addition, EPHA3 overexpression could abolish the inhibitory effects of miR-641 overexpression on the malignant behaviors of glioma cells by activating the signal transducer and activator of transcription 3 (STAT3). These findings elucidated that circ_CLIP2 knockdown suppressed glioma development by regulation of the miR-641/EP HA3/STAT3 axis, which provided a novel mechanism for understanding the pathogenesis of glioma.
大量研究已证实环状RNA（circRNA）是胶质瘤进展中的新型调节因子。在此，我们的工作旨在探究circ_CLIP2在胶质瘤中的具体作用。分别通过qRT-PCR和蛋白质免疫印迹法分析mRNA和蛋白质表达。分别通过MTT法、Transwell法和流式细胞术检测细胞活力、迁移、侵袭和凋亡。此外，通过双荧光素酶报告基因检测和/或RNA免疫沉淀（RIP）检测验证circ_CLIP2、微小RNA（miR）-641和促红细胞生成素产生肝细胞（Eph）A3之间的结合关系。以下数据表明，circ_CLIP2和EPHA3在胶质瘤组织和细胞中显著增加，而miR-647表达下调。功能获得和功能缺失实验发现，circ_CLIP2敲低显著抑制胶质瘤细胞的增殖、迁移和侵袭，并促进其凋亡，而miR-641抑制可消除circ_CLIP2敲低的这些作用。circ_CLIP2被证明是miR-641的海绵，可竞争性上调EPHA3表达。此外,EPHA3过表达可通过激活信号转导和转录激活因子3（STAT3）消除miR-641过表达对胶质瘤细胞恶性行为的抑制作用。这些发现阐明，circ_CLIP2敲低通过调节miR-641/EP HA3/STAT3轴抑制胶质瘤发展，这为理解胶质瘤的发病机制提供了一种新机制。
