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Circ_0084582通过介导miR-485-3p/JAG1轴促进骨肉瘤细胞的生长、迁移、侵袭和血管生成。

Circ_0084582 Facilitates Cell Growth, Migration, Invasion, and Angiopoiesis in Osteosarcoma Mediating the miR-485-3p/JAG1 Axis.

作者信息

Gao Peng, Zhao Xincheng, Yu Keying, Zhu Ziqiang

机构信息

Department of Orthopedics Surgery, The Second Hospital of Xuzhou Coal Mining Group, Xuzhou, China.

Department of Orthopedics Surgery, The General Hospital of Xuzhou Coal Mining Group, Xuzhou, China.

出版信息

Front Genet. 2021 Aug 5;12:690956. doi: 10.3389/fgene.2021.690956. eCollection 2021.

Abstract

Osteosarcoma (OS) is the most representative bone cancer, and circular RNAs serve as pivotal regulators in the progression of OS. This research was designed to explore the role and functional mechanism of circ_0084582 in OS. Circ_0084582, microRNA-485-3p (miR-485-3p), and Jagged1 (JAG1) levels were measured by quantitative real-time polymerase chain reaction. Cell proliferation was examined via 3-(4, 5-dimethylthiazol-2-y1)-2, 5-diphenyl tetrazolium bromide (MTT) assay. Cell cycle progression was analyzed by flow cytometry. Wound healing and transwell assays were performed for evaluating cell migration and invasion. Angiopoiesis was assessed using the tube formation assay. Protein detection was conducted using Western blot. The target relation was identified by the dual-luciferase reporter assay, RNA immunoprecipitation (RIP) assay, and RNA pull-down assay. A xenograft experiment was applied for analyzing the effect of circ_0084582 on OS . Circ_0084582 was highly expressed in OS tissues and cells. Circ_0084582 knockdown reduced cell proliferation, cell cycle progression, migration, invasion, and angiopoiesis of OS cells. JAG1 was upregulated in OS, and its overexpression reversed the effects of circ_0084582 knockdown on OS cells. Circ_0084582 targeted miR-485-3p, and miR-485-3p targeted JAG1, and circ_0084582 could affect the JAG1 level by sponging miR-485-3p. The function of circ_0084582 in OS progression was also achieved by sponging miR-485-3p. Circ_0084582 knockdown decreased OS growth partly by the miR-485-3p-mediated JAG1 downregulation. These results indicate that circ_0084582 functions as a tumorigenic factor in OS via the regulation of miR-485-3p/JAG1 axis.

摘要

骨肉瘤(OS)是最具代表性的骨癌,环状RNA在骨肉瘤进展中起关键调节作用。本研究旨在探讨circ_0084582在骨肉瘤中的作用及功能机制。通过定量实时聚合酶链反应检测circ_0084582、微小RNA-485-3p(miR-485-3p)和锯齿状蛋白1(JAG1)的水平。通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)法检测细胞增殖。通过流式细胞术分析细胞周期进程。进行伤口愈合和Transwell实验以评估细胞迁移和侵袭。使用管腔形成实验评估血管生成。通过蛋白质印迹法进行蛋白质检测。通过双荧光素酶报告基因实验、RNA免疫沉淀(RIP)实验和RNA下拉实验鉴定靶标关系。应用异种移植实验分析circ_0084582对骨肉瘤的影响。circ_0084582在骨肉瘤组织和细胞中高表达。敲低circ_0084582可降低骨肉瘤细胞的增殖、细胞周期进程、迁移、侵袭和血管生成。JAG1在骨肉瘤中上调,其过表达可逆转敲低circ_0084582对骨肉瘤细胞的影响。circ_0084582靶向miR-485-3p,miR-485-3p靶向JAG1,circ_0084582可通过海绵吸附miR-485-3p影响JAG1水平。circ_0084582在骨肉瘤进展中的功能也通过海绵吸附miR-485-3p实现。敲低circ_0084582部分通过miR-485-3p介导的JAG1下调降低骨肉瘤生长。这些结果表明,circ_0084582通过调节miR-485-3p/JAG1轴在骨肉瘤中发挥致瘤因子的作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ff4c/8375504/2e296047a5aa/fgene-12-690956-g001.jpg

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