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Mechanism of METTL3 promoting rituximab resistance via β-catenin/MYC axis in double-expressor diffuse large B cell
Yanming Xue1, Li Miao2, Juan Liu3
1Department of Hematology and Lymphology, Harbin Medical University Cancer Hospital, Harbin, Heilongjiang 150081, People's Republic of China; Department of Hematology, The Second Affiliated Hospital of Harbin Medical University, 246 Xuefu Road, Nangang District, Harbin, Heilongjiang 150001, People's Republic of China.
Objective:
To explore how methyltransferase-like 3 (METTL3) regulates myelocytomatosis oncogene (MYC)/B-cell lymphoma 2 (BCL2) transcriptional expression via YTHDF2-mediated m6A modification of β-catenin, thereby influencing rituximab (RTX) resistance in double-expressor diffuse large B-cell lymphoma (DE-DLBCL).
Methods:
DE-DLBCL cells (U-2932) were cultured in vitro, followed by si-METTL3, oe-β-catenin, oe-MYC, oe-BLC2, or RTX treatment. SRAMP database predicted m6A sites in β-catenin. MeRIP-qPCR and RIP were performed to detect m6A enrichment in β-catenin and YTHDF2 enrichment in β-catenin mRNA. The stability of β-catenin mRNA was assessed using actinomycin D. The interaction between MYC and BCL2 was predicted by JASPAR database and confirmed by dual luciferase assay and ChIP. Finally, a DE-DLBCL nude mouse model was established and treated with RTX and METTL3 inhibitor (STM2457). Tumor growth and related molecule expression were evaluated.
Results:
U-2932 cells exhibited high expression of MYC and BCL2, elevated METTL3 levels, and increased cell viability. Cell cycle progression was accelerated, manifested by decreased cells in the G0-G1 phase, increased cells in S and G2-M phases, elevated cell proliferation, and enhanced RTX resistance. METTL3 knockdown inhibited cell cycle and RTX resistance in DE-DLBCL cells. METTL3 promoted β-catenin expression via m6A modification. METTL3 activated MYC and BCL2 indirectly. METTL3 inhibition suppressed DE-DLBCL cell growth and RTX resistance via the β-catenin/MYC axis in vivo.
Conclusion:
METTL3 may drive DE-DLBCL cell cycle progression and RTX resistance by activating MYC and BCL2 transcription via YTHDF2-mediated m6A modification of β-catenin, revealing a novel mechanism of indirect transcriptional regulation in lymphoma and a potential therapeutic target.
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