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

Peng Gao,Ziqiang Zhu,Keying Yu,Xincheng Zhao

doi:10.3389/fgene.2021.690956

Peng Gao, Ziqiang Zhu + Show 2 more

Open Access

https://doi.org/10.3389/fgene.2021.690956

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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 in vivo. 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 in vivo 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.

Highlights

Osteosarcoma (OS) has become the most common primary bone carcinoma around the world (Lilienthal and Herold, 2020)
The images indicate that linear Glyceraldehyde-3phosphate dehydrogenase (GAPDH) was not detected by divergent primers with cDNA and gDNA as the templates, and circ_0084582 was detected by divergent primers in cDNA (Figure 1D)
It is crucial to research the molecular mechanism during the developing process of OS and seek therapeutic biomarkers for OS treatment

Summary

Introduction

Osteosarcoma (OS) has become the most common primary bone carcinoma around the world (Lilienthal and Herold, 2020). Radiation exposure and alkylating agents are the major risk factors for OS, and rapid bone proliferation is associated with OS (Ritter and Bielack, 2010). It is essential to enhance the understanding of the molecular pathogenetic mechanism of OS and discover novel molecular targets. Circ_0002137 promotes cell invasion and cell cycle progression in OS by sponging miR-433-5p to modulate the IGF1R level (Zhang et al, 2021). Circ_0056285 regulates OS cell apoptosis and proliferation by targeting the miR-1244/TRIM44 axis (Huo and Dou, 2021). Li et al (2020) prove that circ_0000073 contributes to the development of OS by upregulating NRAS via the sponge effects on miR-145-5pa and miR-151-3p. No research focuses on the function and mechanism of circ_0084582 in OS

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