Zhang Peng, Li Jun
Department of Orthopedics, Xiangyang Central Hospital, Affiliated Hospital of Hubei University of Arts and Science, Xiangyang, China.
J Bone Oncol. 2021 Jan 18;27:100349. doi: 10.1016/j.jbo.2021.100349. eCollection 2021 Apr.
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.
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 .
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.
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.
据报道，环状RNA circ_0007534和微小RNA-219a（miR-219a-5p）参与骨肉瘤（OS）的发生发展。骨肉瘤（OS）是最常见的恶性骨肿瘤之一，更容易发生在长骨的干骺端，包括股骨远端和胫骨近端。然而，具体机制尚不完全清楚。本研究的目的是揭示circ_0007534和miR-219a-5p在骨肉瘤中的功能机制。
通过定量实时聚合酶链反应（qRT-PCR）或蛋白质免疫印迹法检测基因水平。采用细胞计数试剂盒-8（CCK-8）和集落形成试验检测细胞增殖能力。使用Transwell试验检测细胞迁移和侵袭能力。此外，通过starbaseV3.0预测miR-219a-5p与circ_0007534或性别决定区Y框蛋白5（SRY）-盒5（SOX5）之间的相互作用，并通过双荧光素酶报告基因试验和RNA免疫沉淀（RIP）试验进行验证。此外，进行肿瘤异种移植实验以分析circ_0007534缺失对肿瘤生长的影响。
在骨肉瘤组织和细胞中，circ_0007534和SOX5水平升高，而miR-219a-5p水平降低。敲低circ_0007534可抑制骨肉瘤细胞的增殖、集落形成、迁移和侵袭。circ_0007534靶向miR-219a-5p，且miR-219a-5p与SOX5相互作用。此外，circ_0007534通过调节miR-219a-5p和SOX5水平来调控骨肉瘤细胞的生长。
我们的研究结果表明，敲低circ_0007534通过调节miR-219a-5p/SOX5轴抑制骨肉瘤细胞生长，为骨肉瘤的治疗和诊断提供了一个潜在靶点。
