Geng Yibo, Wu Yuliang, Xu Cheng, Li Tian, Zhang Liwei
Department of Neurosurgery, Beijing Tiantan Hospital, Capital Medical University, Beijing, People's Republic of China.
Department of Neurosurgery, Qilu Children's Hospital, Shandong University, Jinan, Shandong, People's Republic of China.
Onco Targets Ther. 2020 Oct 9;13:10161-10172. doi: 10.2147/OTT.S272616. eCollection 2020.
Numerous studies suggest that long non-coding RNAs (lncRNAs) participate in the biological process of diverse malignancies, including glioma. Although many differentially expressed lncRNAs have been identified in glioma, to our best knowledge, the role of LINC00662 and its potential underlying mechanism in glioma progression remains unclear. This study aimed to explore the function and regulatory network of LINC00662 in glioma.
Expressions of LINC00662, miR-34a-5p and lectin mannose-binding 2-like (LMAN2L) in glioma tissues were analyzed using The Cancer Genome Atlas Program (TCGA) and the Chinese Glioma Genome Atlas (CGGA) databases. Colony formation, Celltiter-Glo and BrdU (5-bromo-2'-deoxyuridine) incorporation assays were used to detect cell proliferation in vitro. Xenograft mouse models were established to determine cell proliferation in vivo. Transwell and wound healing assay was used to detect cell migration. In addition, epithelial-mesenchymal transition (EMT) markers were detected by Western blot. Annexin V and 7-AAD were used to stain apoptotic cells. Interactions between miR-34a-5p and LINC00662 or the 3'-UTR of LMAN2L were predicted and determined by bioinformatics analysis, luciferase reporter assay and RNA immunoprecipitation (RIP) assays.
High LINC00662 level predicted poor overall survival of glioma patients. Functional studies revealed that suppression of LINC00662 remarkably inhibited cell proliferation, clonogenicity and EMT pathway. Mechanistically, LINC00662 sponged miR-34a-5p to regulate LMAN2L expression. Furthermore, miR-34a-5p inhibitor reversed the anti-proliferation and anti-migration effect of LINC00662 knockdown, which could be rescued by downregulation of LMAN2L in glioma cells.
Our study was the first to report that LINC00662 acted as a competing endogenous RNA (ceRNA) to regulate glioma progression by targeting miR-34a-5p/LMAN2L axis, providing a new therapeutic target for glioma.
大量研究表明，长链非编码RNA（lncRNAs）参与包括胶质瘤在内的多种恶性肿瘤的生物学过程。尽管在胶质瘤中已鉴定出许多差异表达的lncRNAs，但据我们所知，LINC00662在胶质瘤进展中的作用及其潜在机制仍不清楚。本研究旨在探讨LINC00662在胶质瘤中的功能和调控网络。
使用癌症基因组图谱计划（TCGA）和中国胶质瘤基因组图谱（CGGA）数据库分析胶质瘤组织中LINC00662、miR-34a-5p和凝集素甘露糖结合2样蛋白（LMAN2L）的表达。采用集落形成、Celltiter-Glo和BrdU（5-溴-2'-脱氧尿苷）掺入试验检测体外细胞增殖。建立异种移植小鼠模型以确定体内细胞增殖。采用Transwell和伤口愈合试验检测细胞迁移。此外，通过蛋白质免疫印迹法检测上皮-间质转化（EMT）标志物。用膜联蛋白V和7-AAD对凋亡细胞进行染色。通过生物信息学分析、荧光素酶报告基因试验和RNA免疫沉淀（RIP）试验预测并确定miR-34a-5p与LINC00662或LMAN2L的3'-UTR之间的相互作用。
LINC00662水平高预示着胶质瘤患者的总生存期较差。功能研究表明，抑制LINC00662可显著抑制细胞增殖、克隆形成能力和EMT途径。机制上，LINC00662通过吸附miR-34a-5p来调节LMAN2L的表达。此外，miR-34a-5p抑制剂可逆转LINC00662敲低的抗增殖和抗迁移作用，而在胶质瘤细胞中下调LMAN2L可挽救这种作用。
我们的研究首次报道LINC00662作为一种竞争性内源RNA（ceRNA），通过靶向miR-34a-5p/LMAN2L轴调节胶质瘤进展，为胶质瘤提供了一个新的治疗靶点。
