Screening of biomarkers related to early recurrence of hepatocellular carcinoma and their significance in predicting the patients'prognosis

LIANG Meng; CHEN Kehe; CAO Yuhua; PAN Deng; WANG Junping; OU Mei; ZHONG Wenhe; GAO Ting; WEI Haiming

doi:10.3969/j.issn.1674-3806.2025.03.09

Abstract：Objective To screen the biomarkers related to early recurrence of hepatocellular carcinoma(HCC)and to analyze their significance in predicting the HCC patients'prognosis.Methods Based on the previous studies,16 differentially expressed genes(DEGs)were screened,including human leukocyte antigen-A(HLA-A),interleukin-23 alpha(IL-23A),tumor protein 63(TP63),arachidonate 15-lipoxygenase type B(ALOX15B),budding uninhibited by benzimidazoles 1(BUB1),C-X-C motif chemokine receptor 2(CXCR2),C-C motif chemokine ligand 20(CCL20),C-type lectin domain family 4 member C(CLEC4C),protein tyrosine kinase 7(PTK7),myeloperoxidase(MPO),interleukin-1 beta(IL-1β),matrix metallopeptidase 9(MMP9),G antigen 2A(GAGE2A),G antigen 2E(GAGE2E),deleted in malignant brain tumors 1(DMBT1),and forkhead box M1(FOXM1).The Cancer Genome Atlas(TCGA)database was searched with the keyword"TCGA-LIHC",and a total of 374 hepatocellular carcinoma tissues and 50 control tissues(all belonging to cancer-adjacent tissues)were obtained.The RNA databases and their clinical data were downloaded.The expression differences of DEGs in the hepatocellular carcinoma tissues and the cancer-adjacent tissues were analyzed.Gene Ontology(GO)enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were performed on the DEGs.A protein-protein interaction networks(PPI)was constructed by using the STRING online database.The surv_cutpoint function in the R 4.1.2 software package"survminer"was applied to obtain the optimum cut-off value,and the the optimum cut-off value was used to distinguish the high expression group from the low expression group.The relationship between the DEGs and the patients'survival prognosis was analyzed by using Kaplan-Meier survival curves.Cox regression was used to analyze the factors affecting the HCC patients'prognosis.Results Compared with those in the cancer-adjacent tissues,HLA-A,IL-23A,TP63,ALOX15B,BUB1,CCL20,PTK7,MMP9,GAGE2A,DMBT1 and FOXM1 in the hepatocellular carcinoma tissues were highly expressed,while MPO,IL-1β,CXCR2 and CLEC4C in the hepatocellular carcinoma tissues were low expressed,and the differences were statistically significant(P＜0.05).There was no significant difference in the expression of GAGE2E between the hepatocellular carcinoma tissues and the cancer-adjacent tissues(P＞0.05).The results of GO enrichment analysis indicated that the changes in biological process(BP)of DEGs were significantly enriched in neutrophil migration,granulocyte chemotaxis and neutrophil chemotaxis.The changes in cellular components(CC)were predominantly enriched in secretory granule membranes,phagocytic vesicles and phagocytic vesicle membranes.The changes in molecular functions(MF)were enriched in receptor-ligand activity,cytokine receptor binding and cytokine activity.The results of KEGG pathway enrichment analysis revealed that the DEGs were primarily enriched in the tumor necrosis factor(TNF)signaling pathway,interleukin-17(IL-17)signaling pathway and rheumatoid arthritis pathway.The results of PPI analysis showed that the top 5 protein pairs with the highest protein-protein interaction scores were MPO-MMP9(scores:0.969),CCL20-IL-1β(scores:0.958),CCL20-CXCR2(scores:0.949),MMP9-IL-1β(scores:0.949),and IL-1β-MPO(scores:0.860).The results of Kaplan-Meier survival curve analysis indicated that the survival prognoses of the high expression groups of BUB1,MMP9,CCL20,PTK7 IL-1β and FOXM1 were significantly better than those of their corresponding low expression groups(P＜0.05).The results of multivariate Cox regression analysis indicated that T3/T4 stage,tumor-bearing status and high FOXM1 expression were independent risk factors for overall survival(OS)of the HCC patients(P＜0.05).Conclusion DEGs associated with early recurrence of HCC are mainly involved in the pathways related to inflammation and immune responses.FOXM1 may serve as a biological marker for predicting the survival prognosis of the HCC patients.