■环状RNAcirc_0007534和microRNA-219a(miR-219a-5p)被报道参与骨肉瘤(OS)的发展。骨肉瘤(OS)是最常见的恶性骨肿瘤之一。这更容易发生在长骨的干phy端,包括股骨远端和胫骨近端。然而,详细机制尚不完全清楚。本研究旨在揭示circ_0007534和miR-219a-5p在OS中的作用机制。
■通过定量实时聚合酶链反应(qRT-PCR)或蛋白质印迹测定法确定基因的水平。细胞计数试剂盒-8(CCK-8)和集落形成试验检测细胞增殖能力。使用transwell测定法测量细胞迁移和侵袭能力。此外,通过starbaseV3.0预测miR-219a-5p与circ_0007534或SRY(性别决定区Y)-box5(SOX5)之间的相互作用,并通过双荧光素酶报告分析和RNA免疫沉淀(RIP)分析进行证实.此外,进行肿瘤异种移植实验以分析体内circ_0007534耗竭对肿瘤生长的影响。
■circ_0007534和SOX5的水平增加,而OS组织和细胞中miR-219a-5p水平降低。Circ_0007534敲除抑制了扩散,菌落形成,迁移,和在OS细胞中的入侵。Circ_0007534靶向miR-219a-5p,miR-219a-5p与SOX5相互作用。此外,circ_0007534通过调节miR-219a-5p和SOX5的水平来调节OS细胞的生长。
■我们的发现表明,circ_0007534敲低通过调节miR-219a-5p/SOX5轴抑制OS细胞的生长,为OS治疗和诊断提供潜在的目标。
UNASSIGNED: Circular RNA circ_0007534 and microRNA-219a (miR-219a-5p) were reported to be involved in osteosarcoma (OS) development. Osteosarcoma (OS) is one of the most common malignant bone tumors, which was more prone to occur in the metaphysis of long bones, including distal femur and proximal tibia. However, the detailed mechanisms were not fully clear. The purpose of this research was to reveal the functional mechanisms of circ_0007534 and miR-219a-5p in OS.
UNASSIGNED: The levels of genes were determined by quantitative real-time polymerase chain reaction (qRT-PCR) or western blot assay. Cell proliferation ability was detected by cell counting kit-8 (CCK-8) and colony formation assay. Cell migration and invasion abilities were measured using the transwell assay. Furthermore, the interaction between miR-219a-5p and circ_0007534 or SRY (sex-determining region Y)-box 5 (SOX5) was predicted by starbaseV3.0, and confirmed by the dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. Besides, tumor xenograft experiment was performed to analyze the effect of circ_0007534 depletion on tumor growth in vivo.
UNASSIGNED: The levels of circ_0007534 and SOX5 were increased, while the miR-219a-5p level was decreased in OS tissues and cells. Circ_0007534 knockdown repressed the proliferation, colony formation, migration, and invasion in OS cells. Circ_0007534 targeted miR-219a-5p, and miR-219a-5p interacted with SOX5. Furthermore, circ_0007534 regulated the growth of OS cells through modulating the levels of miR-219a-5p and SOX5.
UNASSIGNED: Our finding demonstrated that circ_0007534 knockdown suppressed the growth of OS cells via regulating miR-219a-5p/SOX5 axis, providing a potential target for OS treatment and diagnosis.