mA 读者 YTHDF3 与胃癌的临床预后、相关 RNA 特征和免疫抑制有关。

mA reader YTHDF3 is associated with clinical prognosis, related RNA signatures and immunosuppression in gastric cancer.

机构信息

Department of Gastroenterology, Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200233, China.

Department of Pathology, Zhongshan Hospital, Fudan University, China.

出版信息

Cell Signal. 2023 Aug;108:110699. doi: 10.1016/j.cellsig.2023.110699. Epub 2023 May 5.

DOI:10.1016/j.cellsig.2023.110699

PMID:37149073

Abstract

BACKGROUND

YTHDF3 as a N6-methyladenosine (m6A) reader participates in the development and progression of multiple cancer types, however, the prognosis, molecular biology and immune infiltration of YTHDF3 in gastric cancer (GC) have not been investigated.

METHODS

The YTHDF3 expression profile and clinicopathological parameters of stomach adenocarcinoma (STAD) were downloaded from TCGA. The online websites and databases such as GEPIA2, cBioPortal, UALCAN, ImmuCellAl, xCell, TISIDB, GSCA were utilized for analysis of the association of YTHDF3 with STAD, including clinical prognosis, WGCNA and LASSO Cox regression analysis. Further functional assays such as RT-qPCR, Western blot, immunohistochemistry (IHC), immunofluorescence (IF), CCK-8, colony formation, EdU and Transwell assays were performed to determine the role of YTHDF3 in GC.

RESULTS

We found that YTHDF3 was upregulated in STAD tissue samples ascribed to its copy number amplification and associated with poor prognosis in patients with STAD. GO and KEGG analyses showed that YTHDF3-related differential genes were predominantly enriched in the proliferation, metabolism and immune signaling pathways. Knockdown of YTHDF3 repressed the growth and invasion of GC cells by inhibition of PI3K/AKT signaling. We then identified YTHDF3-related lncRNAs, miRNAs and mRNAs, and constructed their prognostic signatures in patients with STAD. Moreover, YTHDF3 associated with tumor immune infiltration such as CD8+ T cells, macrophages, Tregs, MHC molecules and chemokines, upregulated PD-L1 and CXCL1 and exerted a response to the immunotherapy in GC.

CONCLUSIONS

YTHDF3 upregulation indicates poor prognosis and promotes GC cell growth and invasion by activating PI3K/AKT pathway and regulating immune microenvironment. The established YTHDF3-related signatures highlight the association of YTHDF3 with the clinical prognosis and immune cell infiltration in GC.

摘要

背景

YTHDF3 作为 N6-甲基腺苷（m6A）阅读器，参与多种癌症类型的发生和发展，然而，YTHDF3 在胃癌（GC）中的预后、分子生物学和免疫浸润尚未被研究。

方法

从 TCGA 下载 YTHDF3 表达谱和胃腺癌（STAD）的临床病理参数。利用 GEPIA2、cBioPortal、UALCAN、ImmuCellAl、xCell、TISIDB、GSCA 等在线网站和数据库分析 YTHDF3 与 STAD 的相关性，包括临床预后、WGCNA 和 LASSO Cox 回归分析。进一步通过 RT-qPCR、Western blot、免疫组化（IHC）、免疫荧光（IF）、CCK-8、集落形成、EdU 和 Transwell 实验等功能实验，确定 YTHDF3 在 GC 中的作用。

结果

我们发现 YTHDF3 在 STAD 组织样本中上调，归因于其拷贝数扩增，并与 STAD 患者的预后不良相关。GO 和 KEGG 分析表明，YTHDF3 相关差异基因主要富集在增殖、代谢和免疫信号通路中。敲低 YTHDF3 通过抑制 PI3K/AKT 信号通路抑制 GC 细胞的生长和侵袭。然后，我们鉴定了 YTHDF3 相关的 lncRNA、miRNA 和 mRNA，并构建了它们在 STAD 患者中的预后特征。此外，YTHDF3 与肿瘤免疫浸润相关，如 CD8+T 细胞、巨噬细胞、Tregs、MHC 分子和趋化因子，上调 PD-L1 和 CXCL1，并在 GC 中对免疫治疗有反应。