CD1c作为肝细胞癌复发的潜在生物标志物的鉴定与验证

Identification and validation of CD1c as a potential biomarker for recurrence of hepatocellular carcinoma

目的:构建肝细胞癌复发风险模型,并探究CD1c在肝细胞癌复发中作为生物标志物的潜在价值。方法:从公共数据库TCGA中下载肝细胞癌的临床信息和lncRNA的转录组数据,通过差异分析、Lasso回归分析等生物信息学方法,构建预测肝细胞癌复发的风险模型。然后通过模型给患者进行评分,筛选与肝细胞癌复发有关的免疫基因。最后使用免疫荧光染色对筛选的免疫基因进行验证。结果:肝细胞癌患者肿瘤组织与正常组织共有4539个差异表达的lncRNA(P<0.05),而复发患者与非复发患者肝肿瘤的差异lncRNA共有697个(P<0.05)。二者取交集得到的30个候选lncRNA用于构建预测复发的风险模型。根据风险模型评分公式,将肝细胞癌患者分为高、低复发风险组。在训练组和测试组中,ROC曲线分析显示模型(训练组AUC=0.737,测试组AUC=0.780)相较于其他临床因素具有更好的预测诊断能力。基因集富集分析(GSEA)与基因集变异分析(GSVA)提示肝细胞癌的复发与免疫抑制相关。对风险评分与免疫基因作相关性分析,提示CD1c与肝细胞癌的复发有关(皮尔逊相关系数r=0.291,P<0.001)。免疫荧光染色分析进一步验证了CD1c可以作为生物标志物的能力。结论:基于lncRNA构建的复发风险模型具有优秀的预测肝细胞癌复发的能力,筛选出的免疫基因CD1c可作为肝细胞癌复发的潜在生物标志物。

Objective:To construct a model of hepatocellular carcinoma recurrence risk and explore the potential value of CD1c as a biomarker in hepatocellular carcinoma recurrence.Methods:The clinical information and lncRNA transcriptome data for hepatocellular carcinoma were downloaded from TCGA.Bioinformatics methods such as differential analysis and Lasso regression analysis were used to construct a risk model for predicting the recurrence of hepatocellular carcinoma.After scoring patients by the model,immune genes associated with hepatocellular carcinoma recurrence were screened.Finally,immunofluorescence staining was used to validate the screened immune genes.Results:There were 4539 differentially expressed lncRNAs between the tumor and the normal group,and 697 differentially expressed lncRNAs between the recurrence and the non‐recurrence group.The 30 candidate lncRNAs obtained by the intersection of the above results were used to construct a risk model for predicting recurrence.According to the risk model scoring formula,patients with hepatocellular carcinoma were divided into high and low‐recurrence risk groups.In the training and testing group,ROC curve analysis showed that the model(AUC=0.76 in the training group group and AUC=0.81 in the test group)had better predictive diagnostic ability than other clinical factors.The gene set enrichment analysis(GSEA)and gene set variation analysis(GSVA)suggested that the recurrence of hepatocellular carcinoma was associated with immunosuppression.Correlation analysis between the risk score and immune genes indicated that CD1c was associated with the recurrence of hepatocellular carcinoma.Immunofluorescence staining analysis further confirmed that CD1c could serve as a biomarker.Conclusion:The lncRNA‐based risk model has excellent ability to predict the recurrence of hepatocellular carcinoma and the identified gene CD1c may serve as a potential biomarker for hepatocellular carcinoma recurrence.