Integrative analysis of bulk and single-cell sequencing reveals TNFSF9 as a potential regulator in microsatellite instability stomach adenocarcinoma.

IF 2.8 3区医学 Q2 MEDICINE, RESEARCH & EXPERIMENTAL

European Journal of Medical Research Pub Date : 2025-03-28 DOI:10.1186/s40001-025-02471-0

Jianlong Zhou, Yucheng Zhang, Yongfeng Liu, Jiehui Li, Wenxing Zhang, Junjiang Wang, Xueqing Yao, Huolun Feng, Jiabin Zheng, Yong Li

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引用次数: 0

Abstract

Background: Stomach adenocarcinoma (STAD) with microsatellite instability (MSI) is associated with a better prognosis compared to Non-MSI. This study aims to elucidate the differences in the tumor microenvironment (TME) of MSI and explore its underlying mechanisms in STAD.

Methods: TME differences between MSI and Non-MSI were analyzed using single-cell RNA sequencing (MSI = 7, Non-MSI = 19) and bulk RNA sequencing (MSI = 39, Non-MSI = 198). Differentially expressed genes (DEGs) were used to identify enriched pathways and hub genes. TNFSF9 expression was validated by immunohistochemistry (IHC) on 23 STAD sections (MSI = 13, Non-MSI = 10) and confirmed in tumor epithelial cells using SNU-1 (MSI) and AGS (Non-MSI) cell lines through quantitative polymerase chain reaction (qPCR) and Western blot (WB).

Results: The results showed MSI was significantly associated with a better prognosis (P < 0.05). Within the TME, MSI was associated with a higher abundance of antigen-presenting cells, including M1 macrophages (40.1% vs. 27.9%) and activated dendritic cells (22.1% vs. 10.5%), as well as pro-inflammatory Th1-like CD4⁺ T cells (15% vs. 11%). However, MSI also showed an increase in exhausted T cells, indicating a complex immune landscape. Signaling pathway and cell communication analyses revealed an enrichment of cytokine-related pathways in MSI. Hub gene analysis revealed that TNFSF9 was predominantly expressed in stromal cells and partially in tumor epithelial cells in MSI, with its upregulation further confirmed through IHC, qPCR, and WB. Correlation analysis demonstrated a positive relationship between TNFSF9 expression and the abundance of M1 macrophages.

Conclusions: These findings provide new insights into the TME of MSI in STAD, emphasizing the significant role of TNFSF9 in shaping MSI-specific TME, enhancing immunotherapy efficacy, and improving patient survival.

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整体和单细胞测序的综合分析显示TNFSF9在微卫星不稳定性胃腺癌中是一个潜在的调节因子。

背景：伴有微卫星不稳定性（MSI）的胃腺癌（STAD）与非MSI相比预后更好。本研究旨在阐明MSI肿瘤微环境（tumor microenvironment， TME）的差异，并探讨其在STAD中的潜在机制。方法：采用单细胞RNA测序（MSI = 7, Non-MSI = 19）和批量RNA测序（MSI = 39, Non-MSI = 198）分析MSI与Non-MSI的TME差异。差异表达基因（DEGs）用于鉴定富集通路和枢纽基因。采用免疫组化（IHC）方法在23个STAD切片（MSI = 13, Non-MSI = 10）上验证了TNFSF9的表达，并通过定量聚合酶链反应（qPCR）和Western blot （WB）方法在SNU-1 （MSI）和AGS （Non-MSI）细胞系上证实了TNFSF9在肿瘤上皮细胞中的表达。结论：这些发现为STAD中MSI的TME提供了新的认识，强调了TNFSF9在形成MSI特异性TME、增强免疫治疗疗效、提高患者生存率方面的重要作用。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

European Journal of Medical Research 医学-医学：研究与实验

CiteScore

3.20

自引率

0.00%

发文量

247

审稿时长

>12 weeks

期刊介绍： European Journal of Medical Research publishes translational and clinical research of international interest across all medical disciplines, enabling clinicians and other researchers to learn about developments and innovations within these disciplines and across the boundaries between disciplines. The journal publishes high quality research and reviews and aims to ensure that the results of all well-conducted research are published, regardless of their outcome.