Abstract：
Osteosarcoma (OS) is the most common bone tumour with a high risk of metastatic progression and recurrence after treatment. Circular RNA hsa_circ_0000591 (circ_0000591) plays a compelling role in OS aggressiveness. However, the function and regulatory mechanism of circ_0000591 need to be further elucidated. As a subject of this study, a differential circRNA circ_0000591 was screened by circRNA microarray expression profiling (GSE96964). Expression changes of circ_0000591 were detected using real-time quantitative polymerase chain reaction (RT-qPCR). Effects of circ_0000591 silencing on OS cell viability, proliferation, colony formation, apoptosis, invasion, and glycolysis were determined via functional experiments. The mechanism by which circ_0000591 functions as a molecular sponge for miRNAs was predicted using bioinformatics analysis and validated using dual-luciferase reporter and RNA pull-down assays. Xenograft assay was done to validate the function of circ_0000591. Circ_0000591 was strongly expressed in OS samples and cells. Silencing of circ_0000591 lessened cell viability, repressed cell proliferation, invasion, glycolysis, and promoted cell apoptosis. Importantly, circ_0000591 regulated HK2 expression by serving as a miR-194-5p molecular sponge. MiR-194-5p silencing impaired circ_0000591 downregulation-mediated suppression of OS cell malignancy and glycolysis. HK2 overexpression weakened the inhibiting impacts of miR-194-5p on OS cell malignancy and glycolysis. Also, circ_0000591 silencing decreased xenograft tumour growth in vivo. Circ_0000591 drove OS glycolysis and growth by upregulating HK2 by sequestering miR-194-5p. The study highlighted the tumour-promoting function of circ_0000591 in OS.
摘要：
背景：骨肉瘤（OS）是最常见的骨肿瘤，治疗后转移进展和复发的风险很高。环状RNAhsa_circ_0000591(circ_0000591)在OS侵袭性中起着引人注目的作用。然而,circ_0000591的功能和调控机制有待进一步阐明。
方法：作为本研究的一个主题，通过circRNA微阵列表达谱分析(GSE96964)筛选差异circ_0000591。使用实时定量聚合酶链反应(RT-qPCR)检测circ_0000591的表达变化。circ_0000591沉默对OS细胞活力的影响,扩散,菌落形成,凋亡，入侵,和糖酵解通过功能实验确定。使用生物信息学分析预测circ_0000591充当miRNA的分子海绵的机制，并使用双荧光素酶报告基因和RNA下拉法进行验证。进行异种移植物测定以验证circ_0000591的功能。
结果:Circ_0000591在OS样品和细胞中强烈表达。circ_0000591的沉默降低了细胞活力，抑制细胞增殖，入侵,糖酵解,促进细胞凋亡。重要的是,circ_0000591通过充当miR-194-5p分子海绵来调节HK2表达。MiR-194-5p沉默受损circ_0000591下调介导的OS细胞恶性肿瘤和糖酵解的抑制。HK2过表达削弱了miR-194-5p对OS细胞恶性和糖酵解的抑制作用。此外，circ_0000591沉默降低了体内异种移植肿瘤的生长。
结论：Circ_0000591通过隔离miR-194-5p上调HK2来驱动OS糖酵解和生长。该研究强调了OS中circ_0000591的促肿瘤功能。本文受版权保护。保留所有权利。
方法：作为本研究的一个主题，
结果:Circ_0000591在OS样品和细胞中强烈表达。
结论：Circ_0000591通过隔离miR-194-5p上调HK2来驱动OS糖酵解和生长。
