铜代谢相关基因差异表达对良恶性胸腔积液性质判断的研究

Study on Differential Expression of Copper Metabolism-Related Genes in Differentiating Benign and Malignant Pleural Effusions

摘要: 目的：本研究旨在通过转录组测序分析CMRGs在良恶性胸腔积液中的表达特征，筛选潜在诊断标志物。方法：收集胸腔积液标本进行转录组测序，通过转录组测序及生物信息学分析筛选出目的基因，通过RT-qPCR验证关键基因表达。结果：共筛选出470个差异表达基因，其中411个上调，59个下调。GO/KEGG富集分析显示，差异基因主要与钙离子结合、O-聚糖加工、双细胞紧密连接组件功能等因素相关，在途径方面显著富集于细胞粘附分子、花生四烯酸代谢等通路。与154个铜代谢相关基因取交集，发现显著下调基因MT1E、MT1X和MT2A，ROC曲线显示其对肺癌的诊断效能较高(AUC分别为0.694、0.817和0.840)，其中MT1X和MT2A的诊断效能高于传统指标癌胚抗原。此外，lncRNA LOC105369559与铜代谢相关基因共表达网络关联密切。RT-qPCR验证结果与测序趋势一致，均具有统计学差异。结论：细胞粘附分子途径以及花生四烯酸代谢途径与恶性胸腔积液进展相关；恶性胸腔积液差异表达的lncRNA LOC105369559与铜代谢相关基因关联性最强；差异表达的铜代谢相关基因MT1E、MT1X、MT2A和lncRNA LOC105369559在恶性胸腔积液中的表达显著下调；MT1X和MT2A在引起恶性胸腔积液的肺癌诊断中优于传统指标。

Abstract: Objective: This study aims to analyze the expression characteristics of copper metabolism-related genes (CMRGs) in benign and malignant pleural effusions through transcriptome sequencing and to screen potential diagnostic biomarkers. Methods: Pleural effusion specimens were collected for transcriptome sequencing. Differentially expressed genes (DEGs) were identified using transcriptomic and bioinformatics analyses, followed by RT-qPCR validation of key genes. Results: A total of 470 DEGs were identified, including 411 upregulated and 59 downregulated genes. GO/KEGG enrichment analysis revealed that these DEGs were primarily associated with biological processes such as calcium ion binding, O-glycan processing, and bicellular tight junction assembly. Pathway analysis highlighted significant enrichment in cell adhesion molecules and arachidonic acid metabolism. Intersection with 154 copper metabolism-related genes identified three significantly downregulated genes: MT1E, MT1X, and MT2A. ROC curve analysis demonstrated their high diagnostic efficacy for lung cancer (AUC values: 0.694, 0.817, and 0.840, respectively), with MT1X and MT2A outperforming the traditional biomarker carcinoembryonic antigen (CEA). Additionally, the lncRNA LOC105369559 exhibited a strong co-expression network with copper metabolism-related genes. RT-qPCR validation confirmed consistent expression trends with transcriptome sequencing, showing statistically significant differences. Conclusion: Cell adhesion molecule and arachidonic acid metabolism pathways are implicated in the progression of malignant pleural effusions. The differentially expressed lncRNA LOC105369559 shows the strongest association with copper metabolism-related genes. The copper metabolism-related genes MT1E, MT1X, MT2A, and lncRNA LOC105369559 are significantly downregulated in malignant pleural effusions. Notably, MT1X and MT2A exhibit superior diagnostic performance to CEA in identifying lung cancer-associated malignant pleural effusions.