Background

Osteosarcoma (OS) is a serious bone malignancy that commonly occurred in humans. Recent research suggested that circular RNA (circRNA) Dedicator of cytokinesis 1 (circDOCK1, also called hsa_circ_0020378) enrolled in the tumorigenesis of osteogenic sarcoma. This subject aimed to explore the precise role and mechanism of circDOCK1 on OS progression.

Methods

CircDOCK1, microRNA-936 (miR-936), and Lymphoid enhancer binding factor 1 (LEF1) levels were detected using real-time quantitative polymerase chain reaction (RT-qPCR). Cell Counting Kit-8 (CCK-8), colony formation, 5-ethynyl-2′-deoxyuridine (EdU), transwell, wound healing, and tube formation assays were used to assess OS cell proliferation, migration, invasion, and angiogenesis. Western blot analysis of protein levels of proliferating cell nuclear antigen (PCNA), matrix metalloproteinase 2 (MMP2), MMP9, and LEF1. According to bioinformatics software (circular RNA Interactome and TargetScan) analysis, the binding between miR-936 and circDOCK1 or LEF1 was predicted, followed by verification by a dual-luciferase reporter and RNA Immunoprecipitation (RIP) assays.

Results

Increased circDOCK1 and LEF1, and decreased miR-936 were found in OS tissues and cell lines. Furthermore, circDOCK1 silencing might suppress OS cell proliferation, migration, invasion, and angiogenesis in vitro. Bioinformatics analysis exhibited that circDOCK1 acted as a sponge for miR-936 and LEF1 was a downstream target of miR-936. Moreover, circDOCK1 functions through modulation of the miR-936/LEF1 axis.

Conclusion

CircDOCK1 knockdown might attenuate OS cell malignant biological behaviors by regulating the miR-936/GFRA1 axis, which may highlight the diagnostic and therapeutic potential of these molecules for OS treatment.

中文翻译：

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环状 RNA circDOCK1 通过充当 miR-936 调节 LEF1 的 ceRNA 促进骨肉瘤进展

背景

骨肉瘤（OS）是一种常见于人类的严重骨恶性肿瘤。最近的研究表明，胞质分裂的环状 RNA (circRNA) Dedicator 1 (circDOCK1，也称为 hsa_circ_0020378) 参与了成骨肉瘤的肿瘤发生。本课题旨在探讨circDOCK1在OS进展中的确切作用和机制。

方法

使用实时定量聚合酶链反应 (RT-qPCR) 检测 CircDOCK1、microRNA-936 (miR-936) 和淋巴增强子结合因子 1 (LEF1) 水平。细胞计数 Kit-8 (CCK-8)、集落形成、5-乙炔基-2'-脱氧尿苷 (EdU)、transwell、伤口愈合和管形成测定用于评估 OS 细胞增殖、迁移、侵袭和血管生成。增殖细胞核抗原 (PCNA)、基质金属蛋白酶 2 (MMP2)、MMP9 和 LEF1 的蛋白质水平的蛋白质印迹分析。根据生物信息学软件（circRNA Interactome 和 TargetScan）分析，预测 miR-936 与 circDOCK1 或 LEF1 之间的结合，然后通过双荧光素酶报告基因和 RNA 免疫沉淀 (RIP) 测定进行验证。

结果

在 OS 组织和细胞系中发现 circDOCK1 和 LEF1 增加，miR-936 减少。此外，circDOCK1 沉默可能在体外抑制 OS 细胞增殖、迁移、侵袭和血管生成。生物信息学分析表明，circDOCK1 充当 miR-936 的海绵，LEF1 是 miR-936 的下游靶标。此外，circDOCK1 通过调节 miR-936/LEF1 轴发挥作用。

结论

CircDOCK1 敲低可能通过调节 miR-936/GFRA1 轴来减弱 OS 细胞恶性生物学行为，这可能突出这些分子对 OS 治疗的诊断和治疗潜力。