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METTL3-Mediated Maturation of miR-99a-5p Promotes Cell Migration and Invasion in Oral Squamous Cell Carcinoma by Targeting ZBTB7A

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Abstract

METTL3 is an important methyltransferase in N(6)-methyladenosine (m6A) modification. Recently, METTL3 mediates methylation of pri-microRNA (miRNA) to accelerate miRNA maturation, regulating tumor development. This study explored whether METTL3 mediated miR-99a-5p to influence oral squamous cell carcinoma (OSCC) cell metastasis. MiR-99a-5p, ZBTB7A, and MATTL3 expression was measured using quantitative real-time PCR. Biological behaviors were assessed using cell counting kit-8, flow cytometry, Transwell assay, as well as western blot. Luciferase reporter assay evaluated the interaction between miR-99a-5p and ZBTB7A. METTL3-regulated pri-miR-99a-5p processing was determined by RNA binding protein immunoprecipitation (RIP) and methylated RNA immunoprecipitation (MeRIP) assays. The consequences clarified that miR-99a-5p was upregulated in OSCC cells. Downregulation of miR-99a-5p suppressed cellular viability, migration, invasion, and epithelial-mesenchymal transition (EMT), and induced apoptosis. ZBTB7A acted as a miR-99a-5p target and reversed the effects on cellular behaviors induced by miR-99a-5p inhibitor. m6A content and METTL3 expression were increased in OSCC cells. METTL3 promoted the m6A modification of pri-miR-99a-5p and thereby facilitated miR-99a-5p processing. Moreover, knockdown of METTL3 inhibited OSCC metastasis by downregulating miR-99a-5p. Taken together, METTL3 promoted miR-99a-5p maturation in an m6A-dependent manner, which further targets ZBTB7A to accelerate the progression of OSCC. These findings suggest potential targets for OSCC therapy.

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Funding

This study was supported by Research Project of TCM Bureau of Guangdong Province(Project name: Study on the mechanism of inhibiting invasion and metastasis of oral squamous cell carcinoma by reducing the m6A modification level of miR-99a-5p by downregulating METTL3, Project number: 20222084).

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  1. Department of stomatology, The Second Affiliated Hospital of Guangzhou, University of Chinese Medicine, 6/F, East Zone, No. 111, Dade Road, Yuexiu District, Guangzhou, 510000, Guangdong, China

    Yuhua Huang, Yun Guan & Xing Zhang

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12033_2023_815_MOESM4_ESM.jpg

Supplementary Material 4: Supplementary Figure S1. Effects of miR-99a-5p on the biological functions of HOK cells. After HOK cells were transfected with miR-99a-5p mimic and mimic nc, (A) cell viability was evaluated using CCK-8; (B,C) cell migration and (D,E) invasion were evaluated by Transwell assay.

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Huang, Y., Guan, Y. & Zhang, X. METTL3-Mediated Maturation of miR-99a-5p Promotes Cell Migration and Invasion in Oral Squamous Cell Carcinoma by Targeting ZBTB7A. Mol Biotechnol (2023). https://doi.org/10.1007/s12033-023-00815-x

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