Mesquita Patrícia, Coelho Alexandre, Ribeiro Ana S, Póvoas Luís F C, de Oliveira Catarina, Leça Nelson, Silva Sara, Ferreira Diana, Pádua Diana, Coelho Ricardo, Jerónimo Carmen, Paredes Joana, Conde Carlos, Pereira Bruno, Almeida Raquel
i3S - Institute for Research and Innovation in Health, University of Porto, 4200-135, Porto, Portugal.
IPATIMUP - Institute of Molecular Pathology and Immunology, University of Porto, 4200-465, Porto, Portugal.
Gastric Cancer. 2025 May 14. doi: 10.1007/s10120-025-01620-y.
Gastric cancer (GC) is the fourth most common cause of cancer-related mortality and the fifth most common cancer worldwide. Despite efforts, the identification of biomarkers and new therapeutic approaches for GC remains elusive. Recent studies have begun to reveal the role of N6-adenosine methylation (mA) in the regulation of gene expression.
The expression of the reader YT521-B homology domain-containing family 3 (YTHDF3) in GC was assessed in 331 patients using immunohistochemistry. GC cell lines depleted of YTHDF3 using CRISPR-Cas9 were evaluated for migration, metastasis, orientation of the mitotic spindle, and response to paclitaxel. The association between YTHDF3 and EZRIN (EZR) mRNA was shown using RNA sequencing, immunofluorescence, real-time PCR, and RNA immunoprecipitation. The single-base elongation- and ligation-based qPCR amplification (SELECT) method was used to map mA in the EZR transcript.
YTHDF3 was significantly overexpressed in GC, and high levels of YTHDF3 were predictive of the response to chemotherapy. In GC cell lines, YTHDF3 was the most highly expressed reader protein. YTHDF3 depletion impaired cytoskeleton organization, cell migration and metastasis, and orientation of the mitotic spindle, leading to an increased response to paclitaxel. EZR was one of the downregulated targets in the YTHDF3 knockout cell models and was associated with the observed phenotype.
YTHDF3 contributes to cell motility and response to paclitaxel in GC cell lines, at least in part through EZR regulation. The YTHDF3-EZR regulatory axis is a novel molecular player in GC, with clinical relevance and potential therapeutic utility.
胃癌（GC）是全球癌症相关死亡的第四大常见原因，也是第五大常见癌症。尽管人们做出了努力，但胃癌生物标志物的识别和新的治疗方法仍然难以捉摸。最近的研究开始揭示N6-腺苷甲基化（mA）在基因表达调控中的作用。
使用免疫组织化学方法评估了331例患者中含YT521-B同源结构域家族3（YTHDF3）的阅读蛋白在胃癌中的表达。使用CRISPR-Cas9技术敲除YTHDF3的胃癌细胞系，评估其迁移、转移、有丝分裂纺锤体方向以及对紫杉醇的反应。使用RNA测序、免疫荧光、实时PCR和RNA免疫沉淀技术显示YTHDF3与埃兹蛋白（EZR）mRNA之间的关联。使用基于单碱基延伸和连接的qPCR扩增（SELECT）方法绘制EZR转录本中的mA图谱。
YTHDF3在胃癌中显著过表达，高水平的YTHDF3可预测化疗反应。在胃癌细胞系中，YTHDF3是表达最高的阅读蛋白。YTHDF3的缺失损害了细胞骨架组织、细胞迁移和转移以及有丝分裂纺锤体的方向，导致对紫杉醇的反应增加。EZR是YTHDF3基因敲除细胞模型中下调的靶点之一，并且与观察到的表型相关。
YTHDF3至少部分通过调节EZR促进胃癌细胞系中的细胞运动性和对紫杉醇的反应。YTHDF3-EZR调节轴是胃癌中的一种新型分子机制，具有临床相关性和潜在的治疗用途。
