基于脂质代谢相关转移风险基因的肝细胞癌患者预后预测模型的构建

Construction of a predictive model for the prognosis of patients with hepatocellular carcinoma based on lipid metabolism-related metastasis risk genes

目的基于相关数据库分析筛选肝细胞癌(HCC)脂质代谢相关转移风险基因,并联合其他临床危险因素构建患者的预后预测模型。方法应用R软件从GEO数据库中获得原发性和转移性HCC患者的差异表达基因(DEGs),并筛选与患者预后相关的DEGs。将TCGA数据库中的HCC患者通过层次聚类分为两组,评估两组患者EMT评分、脂质代谢水平和预后。应用ICGC数据库中的数据再次对上述分析进行验证。应用LASSO回归模型筛选脂质代谢相关转移风险基因并进行风险评分,通过风险评分中位数分别将TCGA和ICGC数据库中HCC患者分为高、低危组,并分析患者的预后。应用单因素和多因素Cox回归分析获得影响HCC患者预后的独立危险因素,并构建列线图预后模型。采用Western blot和油红O染色检测应用脂质代谢抑制剂Fatostatin后Huh7细胞脂质代谢的情况;采用qPCR技术检测Huh7细胞中脂质代谢相关转移风险基因表达水平。结果从GEO数据库中获得原发性和转移性HCC患者的DEGs共159个,其中65个DEGs与HCC患者的OS显著相关。通过EMT评分将TCGA数据库中聚类所得的两组HCC患者分别定义为高、低转移风险组。高转移风险组患者脂质代谢评分更高,OS更短。在ICGC数据库中验证的结果与TCGA数据库一致。应用LASSO回归模型筛选出脂质代谢相关转移风险基因,高危组OS更短。将脂质代谢相关转移风险基因与影响HCC患者预后的独立危险因素相结合,构建预后预测列线图模型。细胞实验证实,应用Fatostatin后,Huh7细胞的脂肪酸合酶表达降低,细胞内脂滴含量减少,多种脂质代谢相关转移风险基因表达发生变化。结论基于数据库分析获得了13个脂质代谢相关转移风险基因,将这些基因和临床危险因素联合构建了HCC患者的预后预测模型,并通过细胞实验初步验证了脂质代谢相关转移风险基因与脂质代谢密切相关。

Objective To identify the lipid metabolism-related metastasis risk genes for hepatocellular carcinoma(HCC)based on related databases,and to construct a predictive model for the prognosis of HCC patients in combination with other clinical risk factors.Methods R software was used to obtain the differentially expressed genes(DEGs)between the patients with primary HCC and those with metastatic HCC from the GEO database,and the DEGs associated with the prognosis of patients were identified.The HCC patients in TCGA database were divided into two groups based on hierarchical clustering,and the two groups were assessed in terms of epithelial-mesenchymal transition(EMT),lipid metabolism,and prognosis.The data in the ICGC database were used for validation of the above analysis.The LASSO regression model was used to obtain the lipid metabolism-related metastasis risk genes and determine their risk scores,and according to the median of risk scores,HCC patients in both TCGA and ICGC databases were divided into high and low risk groups to analyze the prognosis of patients.Univariate and multivariate Cox regression analyses were used to obtain independent risk factors for the prognosis of HCC patients,and a nomogram prognostic model was constructed.Western blot and oil red O staining were used to detect the lipid metabolism of Huh7 cells after treatment with the lipid metabolism inhibitor Fatostatin,and qPCR was used to measure the expression levels of lipid metabolism-related metastasis risk genes in Huh7 cells.Results A total of 159 DEGs were obtained from the patients with primary HCC and those with metastatic HCC in the GEO database,among which 65 DEGs were significantly associated with the overall survival(OS)of HCC patients.Based on the EMT score,the two groups of HCC patients obtained by clustering from the TCGA database were defined as high and low metastasis risk groups,respectively,and the patients in the high metastasis risk group tended to have a higher lipid metabolism score and a shorter OS.The validation results in the ICGC database were consistent with the results based on the TCGA database.The LASSO regression model was used to identify the lipid metabolism-related metastasis risk genes,and the high-risk group had a shorter OS.The lipid metabolism-related metastasis risk genes were combined with the independent risk factors for the prognosis of patients with HCC to construct a nomogram prognostic model.Cell experiments confirmed that after the treatment with Fatostatin,there were reductions in the expression of fatty acid synthase and the content of lipid droplets in Huh7 cells,as well as changes in the expression of a variety of lipid metabolism-related metastasis risk genes.Conclusion A total of 13 lipid metabolism-related metastasis risk genes are obtained based on related databases,which are combined with the clinical risk factors to construct a prognostic predictive model for HCC patients,and cell experiments are conducted to confirm that the lipid metabolism-related metastasis risk genes are closely associated with lipid metabolism.