The result of enrich GO analysis.

Cirrhosis is a common endpoint in various chronic liver diseases, and often causes hepatocellular carcinoma. Studies have revealed the significant role of disulfidptosis in the occurrence and development of hepatocellular carcinoma; however, our understanding of this role is limited. Therefore, we aimed to identify potential disulfidptosis-related biomarkers for cirrhosis. We obtained the gene expression data of patients with cirrhosis from the Gene Expression Omnibus (GEO) database. Subsequently, weighted gene co-expression network analysis was performed, and the “limma” package was used to screen for differentially expressed genes (DEGs) associated with disulfidptosis. Significantly altered biological pathways were identified using Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), Gene Set Enrichment Analysis (GSEA), and Gene Set Variation Analysis (GSVA). We constructed protein–protein interaction (PPI) networks using GeneMANIA and generated receiver operating characteristic (ROC) curves to identify hub-shared genes. Additionally, we assessed the distribution of immune cell populations in cirrhotic and control specimens using single-sample GSEA (ssGSEA) and explored their relationship with hub genes. Six hub genes (CXCL12, COL1A1, CXCR4, COL1A2, CCR7, and CXCL8) were closely associated with disulfidptosis-related DEGs. Further immunohistochemical experiments confirmed the potential of CCR7, CXCL12, CXCR4, and CXCL8 as novel diagnostic biomarkers and suggested their potential as new therapeutic targets. These genes mainly promote the development of liver cirrhosis through the oxidative metabolism and cytokine pathways. Furthermore, we observed positive correlations among 23 of the 28 types of immune cells. This study highlights the potential utility of immune cell infiltration and efficient disulfidptosis-related early diagnostic biomarkers in cirrhosis, and highlights its strong useful as a therapeutic target, offering potential clinical application value.

肝硬化是多种慢性肝脏疾病的常见终末阶段，常可诱发肝细胞癌（hepatocellular carcinoma）。已有研究揭示二硫化物细胞死亡（disulfidptosis）在肝细胞癌发生发展中的关键作用，但目前学界对其具体机制的认知仍较为有限。因此本研究旨在筛选与肝硬化相关的潜在二硫化物细胞死亡相关生物标志物。本研究从基因表达综合数据库（Gene Expression Omnibus, GEO）中获取肝硬化患者的基因表达数据，随后开展加权基因共表达网络分析（weighted gene co-expression network analysis），并使用“limma”R包筛选与二硫化物细胞死亡相关的差异表达基因（differentially expressed genes, DEGs）。通过基因本体论（Gene Ontology, GO）、京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes, KEGG）、基因集富集分析（Gene Set Enrichment Analysis, GSEA）及基因集变异分析（Gene Set Variation Analysis, GSVA）识别显著富集的生物学通路。使用GeneMANIA构建蛋白质-蛋白质相互作用（protein–protein interaction, PPI）网络，并生成受试者工作特征（receiver operating characteristic, ROC）曲线以筛选核心共享基因。此外，本研究通过单样本基因集富集分析（single-sample GSEA, ssGSEA）评估肝硬化组织与对照样本的免疫细胞浸润分布情况，并探究其与核心基因的关联。最终得到6个核心基因（CXCL12、COL1A1、CXCR4、COL1A2、CCR7及CXCL8），其与二硫化物细胞死亡相关差异表达基因密切相关。进一步免疫组化实验证实，CCR7、CXCL12、CXCR4及CXCL8有望成为新型诊断生物标志物，并提示其可作为潜在治疗靶点。上述基因主要通过氧化代谢及细胞因子通路促进肝硬化的发生发展。此外，本研究观察到28种免疫细胞中有23种呈现显著正相关。本研究凸显了免疫细胞浸润以及高效的二硫化物细胞死亡相关早期诊断生物标志物在肝硬化诊疗中的应用潜力，同时证实其可作为极具价值的治疗靶点，具备潜在临床转化应用价值。