WTAP-mediated m6A modification of circ_0032463 promotes osteosarcoma progression by sponging miR-145-5p and regulating GFRA1 expression.

Publisher: Wiley Country of Publication: United States NLM ID: 9717231 Publication Model: Print Cited Medium: Internet ISSN: 1099-0461 (Electronic) Linking ISSN: 10956670 NLM ISO Abbreviation: J Biochem Mol Toxicol Subsets: MEDLINE

Original Publication: New York, NY : Wiley, c1998-

MicroRNAs*/genetics ; MicroRNAs*/metabolism ; Osteosarcoma*/genetics ; Osteosarcoma*/pathology ; Osteosarcoma*/metabolism ; RNA, Circular*/genetics ; RNA, Circular*/metabolism ; Glial Cell Line-Derived Neurotrophic Factor Receptors*/genetics ; Glial Cell Line-Derived Neurotrophic Factor Receptors*/metabolism ; Bone Neoplasms*/genetics ; Bone Neoplasms*/metabolism ; Bone Neoplasms*/pathology; Humans ; Animals ; Cell Line, Tumor ; Mice ; Gene Expression Regulation, Neoplastic ; RNA Splicing Factors/genetics ; RNA Splicing Factors/metabolism ; Mice, Nude ; Male ; Mice, Inbred BALB C ; Cell Proliferation/genetics ; Disease Progression ; Female ; RNA, Neoplasm/genetics ; RNA, Neoplasm/metabolism ; Adenosine/analogs & derivatives ; Cell Cycle Proteins

Osteosarcoma (OS) is the most frequent bone malignancy in humans. Previous evidence suggest that circ_0032463 is an oncogenic circular RNA (circRNA) in various cancers, including OS. However, the molecular mechanism of circ_0032463 involved in OS is still unclear. Circ_0032463, microRNA-145-5p (miR-145-5p), GDNF receptor alpha 1 (GFRA1), and Wilms tumor 1-associated protein (WTAP) levels were determined using real-time quantitative polymerase chain reaction (RT-qPCR). Cell proliferation, apoptosis, migration, invasion, and angiogenesis were analyzed using 5-ethynyl-2'-deoxyuridine (EdU), flow cytometry, transwell, and tube formation assays. Western blot analysis was performed to measure matrix metalloproteinase 2 (MMP2), MMP9, GFRA1, and WTAP protein levels. Binding between miR-145-5p and circ_0032463 or GFRA1 was confirmed using a dual-luciferase reporter and pull-down assay. The biological role of circ_0032463 on OS cell growth was also analyzed using a xenograft tumor model in vivo. Methylated RNA immunoprecipitation assay validated the interaction between WTAP and circ_0032463. Circ_0032463, GFRA1, and WTAP levels were increased, and miR-145-5p was decreased in OS tissues and cells. Circ_0032463 deficiency might hinder OS cell proliferation, migration, invasion, angiogenesis, and promote apoptosis in vitro. Mechanically, circ_0032463 worked as a miR-145-5p sponge to increase GFRA1 expression. Repression of circ_0032463 knockdown on tumor cell growth was proved in vivo. Besides, N6-methyladenosine (m6A) modification facilitates the biogenesis of circ_0032463. Taken together, m6A-mediated biogenesis of circ_0032463 facilitates OS cell malignant biological behavior partly via regulating the miR-145-5p/GFRA1 axis, suggesting a promising molecular marker for OS treatment.
(© 2024 Wiley Periodicals LLC.)

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ZDYF2022SHFZ081 Hainan Province Key Research and Development Project

Keywords: GFRA1; WTAP; circ_0032463; miR‐145‐5p; osteosarcoma; proliferation

0 (MicroRNAs)
0 (MIRN145 microRNA, human)
0 (RNA, Circular)
0 (Glial Cell Line-Derived Neurotrophic Factor Receptors)
0 (GFRA1 protein, human)
0 (WTAP protein, human)
0 (RNA Splicing Factors)
CLE6G00625 (N-methyladenosine)
0 (RNA, Neoplasm)
K72T3FS567 (Adenosine)
0 (Cell Cycle Proteins)

Date Created: 20240907 Date Completed: 20240907 Latest Revision: 20241029

20241029

10.1002/jbt.23833

39243199