[Frontiers in Bioscience, Landmark, 25, 1901-1912, June 1, 2020]

MEG3 targets miR-184 and Wnt/β-catenin and modulates properties of osteosarcoma

Liang Li1, SiBiao Pei2, Na Sun3

1Department of Orthopaedics, Nanchong central hospital, Sichuan province, 637000, PR China, 2Department of pharmacy, Nanchong central hospital, Sichuan province, 637000, PR China, 3Department of endocrinology department, Nanchong central hospital, Sichuan province, 637000, PR China

Table of contents

1. Abstract
2. Introduction
3. Materials and methods
    3.1. Cell lines, clinical samples and animals
    3.2. Cell transfection and MEG3 luciferase reporter assay
    3.3. Osteosarcoma model and groups
    3.4. RT-PCR and quantitative real-time PCR (qPCR)
    3.5. CCK-8 assay
    3.6. Analysis of apoptosis
    3.7. Transwell mobility assay
    3.8. Western bloting
    3.9. TOP/FOP-FLASH luciferase reporter assay
    3.10. Immunohistochemical staining
    3.11. Statistical analysis
4. Results
    4.1. Expression of MEG3 and miR-184 in OS
    4.2. MEG3 regulates the OS tumor cell properties by targeting miR-184
    4.3. MEG3 represses the Wnt/β-catenin pathway via targeting miR-184
    4.4. MEG3 inhibits the OS by targeting miR-184 in vivo
5. Discussion
6. References

1. ABSTRACT

Recent studies indicate that MEG3, a long non-coding RNA (lncRNA) and microRNA-184 (miR-184) are abnormally expressed in osteosarcoma (OS). To this end, we show here that MEG3 negatively regulates the expression of miR-184 and down-stream effectors of WNT/β-catenin pathway including β-catenin, T-cell factor 4 (TCF4) and c-MYC. MEG3 overexpression by adenoviral vectors down-regulate proliferation, migration and apoptosis of OS in vitro and restrict the tumor growth in vivo. We also show that the effects of MEG3 can be effectively reversed by miR-184 mimic. Together these studies show that both MEG3 and miR-184 cooperatively regulate the proliferation, migration and apoptosis of OS.

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Abbreviations: lncRNA: long non-coding RNA; MEG3: Maternally Expressed Gene 3; OS: osteosarcoma; miRNA: MicroRNA; VEGF: vascular endothelial growth factor; TCF4: T-cell factor 4; FITC: fluorescein isothiocyanate; PCNA: Protein levels of proliferating cell nuclear antigen; BCL-2: B-cell lymphoma 2; MMP9: matrix metalloproteinase 9; NC mimics: negative control mimics; MEG3 wt, MEG3 wild type; MEG3 mut, MEG3 mutant.

Key Words: MEG3, Osteosarcoma, miR-184, Wnt/β-catenin

Send correspondence to: Liang Li, Nanchong central hospital. 97 Renmin South Road, Shunqing district, Nanchong, Sichuan, PR China, Tel: 86-027-67812888, Fax: 86-027-67812888, E-mail: v2s5w0@sina.com