目的:通过生物信息学技术分析影响肝癌患者生存预后的铁死亡调控基因(ferroptosis-related genes，FRGs)，并在细胞和组织层面进行验证。方法:从TCGA数据库下载肝癌基因表达矩阵和临床数据集，并从ferrdb数据库获取与铁死亡相关的基因。使用LASSO回归分析，构建风险比例模型，并通过受试者工作特征曲线(receiver operating characteristic，ROC)回顾患者1、2、3年生存率，使用单因素和多因素COX回归分析筛选肝癌的独立预后因素。并对得到的基因进行基因本体(Gene ontology，GO)和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes，KEGG)富集分析。最终进行免疫细胞和免疫功能相关评分。在人类蛋白质图谱数据库下载肝癌预后相关的FRGs的正常肝组织和肝癌组织的免疫组化的结果图。并在正常肝细胞L02和肝癌细胞HepG2，临床获取的6名肝癌患者手术切除的肝癌组织和癌旁组织，通过qPCR分析比较TOP20基因的表达量。结果:通过分析共得到了42个影响肝癌患者预后的相关差异表达FRGs。通过LASSO回归分析，最终筛选出11个FRGs(G6PD、HRAS、SLC1A5等)来构建肝癌患者的风险模型。生存分析发现高风险组患者的生存率显著较低风险组小，且高风险组的患者的免疫评分显著高于低风险组。肿瘤的分期和本研究发现的风险因素评分可以作为肝癌患者的独立预后因素。富集分析发现相关FRGs主要涉及细胞周期、脂质氧化等生物学功能。结论:FRGs能够影响肝癌患者的预后，可能是通过参与调节肿瘤细胞的氧化应激和免疫微环境实现的。
Screening of ferroptosis-related genes affecting the prognosis of hepatocellular carcinoma patients based on bioinformatics analysis and experimental validation
Objective:To screen ferroptosis-related genes(FRGs)affecting the prognosis of hepatocellular carcinoma patients based on bioinformatics technology.and validate them at cellular and tissue levels.Methods:Gene expression and clinical datasets of hepatocellular carcinoma were downloaded from TCGA database,and iron death-related genes were obtained from ferrdb data-base.LASSO regression analysis was used to construct a risk-proportional model and to compare patients'1-,2-,and 3-year sur-vival rates by receiver operating characteristic curves(ROC),and independent prognostic factors for hepatocellular carcinoma were screened using univariate and multivariate COX survival analyses.Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were performed for FRGs associated with liver cancer prognosis.Final immune cell and immune function-related scores were performed.The resultant maps of immunohistochemistry of normal liver tissues and hepa-tocellular carcinoma tissues were downloaded from the Human Gene Protein Data Bank for hepatocellular carcinoma prognosis re-lated FRGs.Moreover the expression of TOP20 gene was compared by qPCR analysis in normal hepatocyte L02 and hepatocellu-lar carcinoma cell HepG2,clinically obtained hepatocellular carcinoma tissues and Para cancerous tissues of six hepatocellular carci-noma patients surgically resected.Results:A total of 42 differential FRGs affecting the prognosis of hepatocellular carcinoma pa-tients were obtained by analysis.eleven FRGs(G6PD,HRAS,SLC1A5,etc.)were finally screened by LASSO regression anal-ysis to construct a risk model for hepatocellular carcinoma patients.The survival analysis revealed that the survival rate of patients in the high-risk group was significantly lower than that of the low-risk group,and the immune score of patients in the high-risk group was significantly higher than that of the low-risk group.Tumor stage and risk value can be used as independent prognostic factors for hepatocellular carcinoma patients.Enrichment analysis revealed that relevant FRGs were mainly involved in biological functions such as cell cycle and lipid oxidation.Conclusion:FRGs can influence the prognosis of hepatocellular carcinoma pa-tients,which may be achieved by participating in the regulation of oxidative metabolism and immune microenvironment of tumor cells.
国家科技期刊平台
NSTL
万方数据
维普
