Department of Orthopedics, Jingzhou Hospital Affiliated to Yangtze University, Jingzhou, 434000, China.
Department of Orthopedics, Gong An Country People's Hospital, Jingzhou, 434050, China.
Funct Integr Genomics. 2023 Aug 8;23(3):270. doi: 10.1007/s10142-023-01197-8.
Even though circular RNAs (circRNAs), a class of non-coding endogenous RNA, play a crucial role in the progression of osteosarcoma (OS), the specific function of hsa_circ_0000028 (circUSP48) remains unclear. This study aims to elucidate the mechanism by which circUSP48 regulates OS. We employed qRT-PCR and western blot techniques to quantify circDOCK1, miR-186, and DNMT3A levels. Cell proliferation was assessed using the cell counting kit-8 (CCK-8), 5-Ethynyl-20-deoxyuridine (EdU) assay, and colony formation assay. Cell migration and invasion were evaluated through Transwell and cell scratch assays. Furthermore, we performed dual-luciferase reporter, RIP, and RNA pull-down assays to investigate the association between circUSP48, miR-365, and CYR61. In addition, an in vivo xenograft model was utilized to assess the functional role of circUSP48. High levels of circUSP48 and CYR61 were observed in OS tissues and cells, while miR-365 levels were low. Knockdown of circUSP48 suppressed the multiplication, motility, and invasion of OS cells, thereby reducing carcinoma growth. Moreover, inhibition of miR-365 reversed the OS cell-suppressive effect caused by circUSP48 knockdown through direct interaction with circUSP48. Additionally, circUSP48 upregulated the expression of CYR61 by sponging miR-365. The findings suggest that circUSP48 promotes malignant behavior in OS by regulating the expression of CYR61 through miR-365, making it a potential therapeutic target for OS.
尽管环状 RNA(circRNAs)作为一类非编码内源性 RNA,在骨肉瘤(OS)的进展中发挥着关键作用,但 hsa_circ_0000028(circUSP48)的具体功能仍不清楚。本研究旨在阐明 circUSP48 调节 OS 的机制。我们采用 qRT-PCR 和 Western blot 技术来量化 circDOCK1、miR-186 和 DNMT3A 的水平。通过细胞计数试剂盒-8(CCK-8)、5-乙炔基-20-脱氧尿苷(EdU)检测和集落形成检测来评估细胞增殖。通过 Transwell 和细胞划痕实验评估细胞迁移和侵袭。此外,我们进行了双荧光素酶报告、RIP 和 RNA 下拉实验,以研究 circUSP48、miR-365 和 CYR61 之间的关联。此外,还利用体内异种移植模型来评估 circUSP48 的功能作用。在 OS 组织和细胞中观察到 circUSP48 和 CYR61 水平升高,而 miR-365 水平降低。circUSP48 的敲低抑制了 OS 细胞的增殖、迁移和侵袭,从而减少了癌的生长。此外,通过与 circUSP48 直接相互作用,抑制 miR-365 逆转了 circUSP48 敲低引起的 OS 细胞抑制作用。此外,circUSP48 通过海绵吸附 miR-365 上调了 CYR61 的表达。这些发现表明,circUSP48 通过调节 miR-365 表达来促进 OS 中的恶性行为,使其成为 OS 的潜在治疗靶点。