YTHDF2 promotes gallbladder cancer progression and gemcitabine resistance via m6A-dependent DAPK3 degradation

Sep 24, 2026

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N6-methyladenosine (m6A) is the most abundant internal eukaryotic RNA modification and is implicated in carcinogenesis, yet its roles in gallbladder cancer (GBC) remain unclear. This study examines YTHDF2, an m6A reader, in GBC and finds marked upregulation in tumor tissues. YTHDF2 overexpression enhances GBC proliferation, tumor growth, migration, and invasion while suppressing apoptosis in vitro and in vivo; knockdown shows opposite effects. Mechanistically, YTHDF2 directly targets DAPK3 mRNA via binding to its 3′-UTR and promotes m6A-dependent degradation. DAPK3 inhibition reverses the tumor-suppressive phenotype caused by YTHDF2 loss, and the YTHDF2/DAPK3 axis induces gemcitabine resistance, suggesting therapeutic opportunities.

LanguageEnglish

CategoryHealthcare

FormatPDF

Pages15

Reading Time38 min

AccessFree to download

Abstract

Keywords

Introduction

Background on gallbladder carcinoma and outcomes

m6A RNA modification and YTHDF2 relevance

Received: 2 May 2023 | Revised: 20 August 2023 | Accepted: 23 August 2023 DOI: 10. 1111/cas.15953 OR I G I NAL ART I C L E YTHDF2 promotes gallbladder cancer progression and gemcitabine resistance via m6A-dependent DAPK3 degradation Xuesong Bai1 | Jiemin Chen2 | Wenqin Zhang2 | Shengnan Zhou1 | Liangbo Dong1 | Jianhao Huang1 | Xiaodong He1 1Department of General Surgery, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Peking Union Medical College & Chinese Academy of Medical Sciences, Beijing, China 2Department of Gastroenterology, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Peking Union Medical College & Chinese Academy of Medical Sciences, Beijing, China Correspondence Xiaodong He, Department of General Surgery, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Peking Union Medical College & Chinese Academy of Medical Sciences, No. 1 Shuaifuyuan, Dongcheng District, Beijing 100730, China. Email: [hxdpumch@163.com](hxdpumch@163.com) Funding information Chinese Academy of Medical Sciences Initiative for Innovative Medicine, Grant/ Award Number: 2021-I2M-1-022 Abstract N6-methyladenosine (m6A) is the most abundant internal modification in eukaryotic RNA and involved in the carcinogenesis of various malignancies. However, the functions and mechanisms of m6A in gallbladder cancer (GBC) remain unclear. In this study, we investigated the role and underlying mechanism of the RNA-binding protein YT521-B homology domain-containing family protein 2 (YTHDF2), an m6A reader, in GBC. Herein, we detected that YTHDF2 was remarkably upregulated in GBC tissues compared to normal gallbladder tissues. Functionally, YTHDF2 overexpression promoted the proliferation, tumor growth, migration, and invasion of GBC cells while inhibiting the apoptosis in vitro and in vivo. Conversely, YTHDF2 knockdown induced opposite results. Mechanistically, we further investigated the underlying mechanism by integrating RNA immunoprecipitation sequencing (RIP-seq), m6A-modified RIP-seq, and RNA sequencing, which revealed that death-associated protein kinase 3 (DAPK3) is a direct target of YTHDF2 . YTHDF2 binds to the 3′-UTR of DAPK3 mRNA and facilitates its degradation in an m6A-dependent manner. DAPK3 inhibition restores the tumor-suppressive phenotype induced by YTHDF2 deficiency. Moreover, the YTHDF2/DAPK3 axis induces the resistance of GBC cells to gemcitabine. In conclusion, we reveal the oncogenic role of YTHDF2 in GBC, demonstrating that YTHDF2 increases the mRNA degradation of the tumor suppressor DAPK3 in an m6A-dependent way, which promotes GBC progression and desensitizes GBC cells to gemcitabine. Our findings provide novel insights into potential therapeutic strategies for GBC. K E Y WO R D S cancer progression, DAPK3, gallbladder cancer, m6A modification, YTHDF2 Abbreviations: Act-D, actinomycin D; CNOT1, CCR4-NOT transcription complex subunit 1; co-IP, coimmunoprecipitation; DAPK3, death-associated protein kinase 3; DEG, differentially expressed gene; GBC, gallbladder cancer; GO, Gene Ontology; GSEA, Gene Set Enrichment Analysis; IHC, immunohistochemistry; KEGG, Kyoto Encyclopedia of Genes and Genomes; m6A, N6-methyladenosine; MeRIP-seq, m6A-modified RNA immunoprecipitation and sequencing; Mut, mutant; RIP-seq, RNA immunoprecipitation sequencing; RNA-seq, RNA sequencing; RT-qPCR, real-time quantitative PCR; TIMER, Tumor Immune Estimation Resource online; YTHDF2, YT521-B homology domain-containing family proteins 2. Xuesong Bai, Jiemin Chen, and Wenqin Zhang contributed equally to this work and share first authorship. This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modificat

FAQs

What is the main focus of this study in gallbladder cancer?

The study investigates how the m6A reader YTHDF2 regulates gallbladder cancer progression and affects sensitivity to gemcitabine through its downstream targets.

How does YTHDF2 affect GBC cell behavior in the study?

YTHDF2 overexpression promotes GBC proliferation, tumor growth, migration, and invasion and inhibits apoptosis, while YTHDF2 knockdown produces opposite effects both in vitro and in vivo.

What molecular mechanism links YTHDF2 to gemcitabine resistance?

YTHDF2 binds the 3′-UTR of DAPK3 mRNA and facilitates its m6A-dependent degradation. DAPK3 inhibition restores the tumor-promoting and drug-resistance phenotypes associated with YTHDF2 activity.

Sep 24, 2026

Uploaded by Elsa