上海交通大学医学院附属仁济医院-胃肠病学

多组学整合分析识别胃癌预后标志物MAGEA3、VWCE及其肿瘤微环境调控作用

作者：王文波, 赵永奎

单位：内蒙古医科大学附属赤峰市医院

基金项目：

摘要：

背景：胃癌是全球最常见的消化道恶性肿瘤之一，发展受多因素驱动，目前有效的分子标志物仍较有限，制约了精准预后评估和个体化治疗的临床需求。目的：通过整合多组学数据，识别与胃癌发生、发展和预后相关的关键基因，并探讨其在肿瘤微环境中的表达特征。方法：基于TCGA-STAD队列的转录组数据，进行差异表达基因（DEGs）分析和加权基因共表达网络分析（WGCNA），取交集获得候选关键基因，随后对交集基因进行GO和KEGG富集分析。进一步通过单因素Cox回归和LASSO回归分析筛选预后相关基因，并利用Kaplan-Meier法验证其预后评估价值。联合GEO单细胞数据集GSE167297，分析关键基因在胃癌微环境各细胞亚群中的表达分布。培养人胃癌细胞株HGC-27和AGS，分别构建MAGEA3过表达和VWCE敲低的稳转细胞株，以实时定量PCR法检测MAGEA3和VWCE mRNA表达水平，CCK-8法检测细胞增殖，流式细胞术检测细胞凋亡情况。结果：WGCNA识别出与胃癌组织显著正相关的MEblue模块；共筛选出4 514个DEGs，两者交集得到511个候选基因，富集分析显示这些基因参与细胞周期、DNA复制、p53信号通路等生物学过程。单因素Cox-LASSO回归分析筛选出14个与总生存显著相关的基因，其中MAGEA3和VWCE高表达组提示患者预后不良（P<0.05）。单细胞分析进一步揭示MAGEA3和VWCE在肿瘤相关成纤维细胞中高表达。在HGC-27细胞中，与阴性对照组相比，过表达MAGEA3组的mRNA表达水平显著上调（P均<0.01），细胞活力显著增加（P均<0.01），同时细胞凋亡受到抑制；而在AGS细胞中，与阴性对照组相比，敲低VWCE组的mRNA表达水平显著下调（P均<0.01），细胞活力显著降低（P均<0.001），且细胞凋亡增加。结论：MAGEA3和VWCE可能作为胃癌潜在的预后生物学标志物。

关键词：胃肿瘤；生存分析；差异表达基因；WGCNA；MAGEA3；VWCE

Identification of MAGEA3 and VWCE as Prognostic Biomarkers in Gastric Cancer and Their Roles in Tumor Microenvironment Based on Multi-omics Integration Analysis

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Abstract:

Background: Gastric cancer is one of the most common malignant tumors of the digestive system worldwide. Its development is driven by multiple factors. However, effective molecular biomarkers remain limited, which constrains accurate prognosis assessment and personalized treatment. Aims: To identify key genes associated with the initiation, progression, and prognosis of gastric cancer by integrating multi-omics data, and to explore their expression characteristics within the tumor microenvironment. Methods: Based on transcriptomic data from the TCGA-STAD cohort, differentially expressed genes (DEGs) were identified and a weighted gene co-expression network analysis (WGCNA) was performed. The intersection of the results was taken to obtain candidate key genes, which were then subjected to GO and KEGG enrichment analyses. Prognosis-related genes were further screened using univariate Cox regression and LASSO regression analyses, and their prognostic values were validated using Kaplan-Meier method. The single-cell dataset GSE167297 from GEO was utilized to analyze the expression distribution of key genes within cellular subpopulations of the gastric cancer microenvironment. Human gastric cancer cell lines HGC-27 and AGS were cultured, and stable cell lines with MAGEA3 overexpression and VWCE knockdown were constructed, respectively. Real-time quantitative PCR was used to detect the mRNA expression levels of MAGEA3 and VWCE. Cell proliferation was assessed using CCK-8 assay, and cell apoptosis was examined via flow cytometry. Results: WGCNA identified the MEblue module as being significantly positively correlated with gastric cancer tissue. A total of 4 514 DEGs were screened, and their intersection with the module genes yielded 511 candidate genes. Enrichment analysis indicated their involvement in processes such as the cell cycle, DNA replication, and p53 signaling pathway. Univariate Cox-LASSO regression analyses identified 14 genes significantly associated with overall survival. Among these, high expressions of MAGEA3 and VWCE predicted poor prognosis in patients (P<0.05). Single-cell analysis further revealed high expressions of MAGEA3 and VWCE in cancer-associated fibroblasts. In HGC-27 cells, compared to the negative control group, the overexpression group showed significantly upregulated MAGEA3 mRNA expression level (all P<0.01), significantly increased cell viability (all P<0.01), and inhibited apoptosis. In AGS cells, compared to the negative control group, the knockdown group exhibited significantly downregulated VWCE mRNA expression level (all P<0.01), significantly reduced cell viability at all time points (all P<0.001), and increased apoptosis. Conclusions: MAGEA3 and VWCE may serve as potential prognostic biomarkers for gastric cancer.

Keywords: Stomach Neoplasms; Survival Analysis; Differentially Expressed Genes; WGCNA; MAGEA3; VWCE

投稿时间：2025-10-28