Osteosarcoma is a prevalent malignant bone cancer. This study aimed to explore the biologic role and potential mechanism of circ_0003732 in osteosarcoma carcinogenesis. Quantitative real-time PCR was implemented to detect the RNA expression of circ_0003732, microRNA-377-3p (miR-377-3p) and cytoplasmic polyadenylation element-binding protein 1 (CPEB1). Cell proliferation was evaluated by cell counting kit-8 assay and colony formation assay. Transwell, wound healing and flow cytometry assays were employed to assess cell migration, invasion and apoptosis. In addition, the interaction between miR-377-3p and circ_0003732 or CPEB1 was validated by dual-luciferase reporter assay. The protein expression was detected by western blot assay or immunohistochemistry assay. Xenograft tumor assay was performed to explore the regulation of circ_0003732 on osteosarcoma tumor growth in vivo. Circ_0003732 was upregulated in osteosarcoma tissues and cells. Knockdown of circ_0003732 suppressed osteosarcoma cell proliferation, migration, invasion and triggered cell apoptosis in vitro, as well as reduced osteosarcoma tumor growth in vivo. Meanwhile, miR-377-3p could bind to circ_0003732 and CPEB1 and miR-377-3p inhibitor could reverse the effects of circ_0003732 silence on osteosarcoma cell progression. Furthermore, CPEB1 overexpression could overturn the suppressive impacts of miR-377-3p on osteosarcoma progression. In addition, circ_0003732 silence restrained Wnt/β-catenin signaling pathway via regulating miR-377-3p in osteosarcoma cells. Circ_0003732 might play a positive role in the malignant progression of osteosarcoma by regulating the miR-377-3p/CPEB1 axis and activating the Wnt/β-catenin signaling pathway, which might provide new insights for osteosarcoma therapy.

中文翻译：

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Circ_0003732通过调节miR-377-3p/CPEB1轴和Wnt/β-catenin信号通路促进骨肉瘤进展。

骨肉瘤是一种常见的恶性骨癌。本研究旨在探讨circ_0003732在骨肉瘤癌变中的生物学作用和潜在机制。采用实时定量PCR检测circ_0003732、microRNA-377-3p（miR-377-3p）和细胞质多腺苷酸化元件结合蛋白1（CPEB1）的RNA表达。通过细胞计数试剂盒8测定和集落形成测定评估细胞增殖。采用 Transwell、伤口愈合和流式细胞术测定来评估细胞迁移、侵袭和凋亡。此外，通过双荧光素酶报告基因测定验证了 miR-377-3p 与 circ_0003732 或 CPEB1 之间的相互作用。通过蛋白质印迹法或免疫组化法检测蛋白表达。进行异种移植肿瘤测定以探索circ_0003732对体内骨肉瘤肿瘤生长的调节。Circ_0003732 在骨肉瘤组织和细胞中表达上调。circ_0003732的敲低可抑制体外骨肉瘤细胞的增殖、迁移、侵袭并引发细胞凋亡，并减少体内骨肉瘤肿瘤的生长。同时，miR-377-3p可以与circ_0003732结合，CPEB1和miR-377-3p抑制剂可以逆转circ_0003732沉默对骨肉瘤细胞进展的影响。此外，CPEB1 过表达可以推翻 miR-377-3p 对骨肉瘤进展的抑制作用。此外，circ_0003732沉默通过调节骨肉瘤细胞中的miR-377-3p抑制Wnt/β-catenin信号通路。Circ_0003732可能通过调节miR-377-3p/CPEB1轴、激活Wnt/β-catenin信号通路在骨肉瘤的恶性进展中发挥积极作用，这可能为骨肉瘤的治疗提供新的见解。