目的:通过生物信息学方法分析原位和淋巴结转移肝癌中巨噬细胞异质性标志基因及生存期分析。 方法:从美国国家生物技术信息中心GEO数据库下载肝癌相关单细胞测序数据，GSE编号为GSE149614，通过单细胞聚类、差异基因表达分析获得原位和淋巴结转移肝癌巨噬细胞异质性标志基因，使用在线分析网站STRING进行蛋白质互作网络分析，并对差异表达的Top基因进行GO聚类和GSEA分析，并在GEPIA网站进行生存期分析。 结果:共筛选出70个差异基因，其中50个差异基因显著下调（‑FC=2.26~7.93），20个差异基因显著上调（FC=1.52~2.99）。GO功能注释富集分析发现，原位肝癌巨噬细胞主要为蛋白质异常折叠，而淋巴结转移肝癌巨噬细胞主要为创伤愈合、胞内蛋白质合成、丙酮酸盐（酯）代谢过程。通过生存期分析发现，淋巴结转移肝癌巨噬细胞的5个的标志基因（ALOX5AP、CD109、MMP12、SLC16A10、SPINK1）中MMP12和SPINK1基因低表达时患者的生存期较长（P<0.05）。 结论:原位和淋巴结转移肝癌中巨噬细胞异质性基因与患者的生存期存在显著相关性。

Objective:To analyze macrophage heterogeneity marker genes in hepatocellular carcinoma （HCC） in situ and lymph node tissues and to analyze the overall survival （OS） by bioinformatics methods. Methods:The single‑cell sequencing data related to HCC were downloaded from the GEO database of National Center for Biotechnology Information （GSE number： GSE149614）. Obtaining heterogeneity marker genes of macrophages in primary and lymph node metastatic liver cancer through single-cell clustering and differential gene expression analysis.The online analysis website STRING was used for protein‑protein interaction network analysis， and the differentially expressed Top genes were analyzed by GO clustering and GSEA analysis. The OS analysis was performed on GEPIA website. Results:A total of 70 differentially expressed genes were screened out， of which 50 differentially expressed genes were significantly down‑regulated （‑FC=2.26‑7.93） and 20 differentially expressed genes were significantly up‑regulated （FC=1.52‑2.99）. GO functional annotation enrichment analysis showed that macrophages in situ were mainly involved in abnormal protein folding， while macrophages in lymph nodes were mainly involved in wound healing， intracellular protein synthesis， and pyruvate （ester） metabolism. The results of OS analysis showed that low expression of MMP12 and SPINK1 in 5 marker genes （ALOX5AP， CD109， MMP12， SLC16A10， SPINK1） of macrophages in metastatic lymphoid tissue was associated with a longer survival time （P<0.05）. Conclusion:Macrophage heterogeneity genes in primary and lymph node metastatic HCC are significantly correlated with the OS of patients.

R96

三亚市人才开发专项资金项目［（2023）14?04］