Table_1_Integrated multi-omics analysis identifies ENY2 as a predictor of recurrence and a regulator of telomere maintenance in hepatocellular carcinoma.docx

Hepatocellular carcinoma (HCC) is the most common type of primary liver cancer and has a high recurrence rate. Accurate prediction of recurrence risk is urgently required for tailoring personalized treatment programs for individual HCC patients in advance. In this study, we analyzed a gene expression dataset from an HCC cohort with 247 samples and identified five genes including ENY2, GPAA1, NDUFA4L2, NEDD9, and NRP1 as the variables for the prediction of HCC recurrence, especially the early recurrence. The Cox model and risks score were validated in two public HCC cohorts (GSE76427 and The Cancer Genome Atlas (TCGA)) and one cohort from Huashan Hospital, which included a total of 641 samples. Moreover, the multivariate Cox regression analysis revealed that the risk score could serve as an independent prognostic factor in the prediction of HCC recurrence. In addition, we found that ENY2, GPAA1, and NDUFA4L2 were significantly upregulated in HCC of the two validation cohorts, and ENY2 had significantly higher expression levels than another four genes in malignant cells, suggesting that ENY2 might play key roles in malignant cells. The cell line analysis revealed that ENY2 could promote cell cycle progression, cell proliferation, migration, and invasion. The functional analysis of the genes correlated with ENY2 revealed that ENY2 might be involved in telomere maintenance, one of the fundamental hallmarks of cancer. In conclusion, our data indicate that ENY2 may regulate the malignant phenotypes of HCC via activating telomere maintenance.

肝细胞癌（Hepatocellular carcinoma, HCC）是原发性肝癌最常见的病理类型，且具有较高的复发率。精准预测复发风险是临床亟需，可为提前为肝细胞癌患者制定个体化诊疗方案提供关键依据。本研究分析了一项包含247例样本的肝细胞癌队列基因表达数据集，筛选出ENY2、GPAA1、NDUFA4L2、NEDD9及NRP1共5个基因，作为肝细胞癌复发（尤其是早期复发）的预测特征变量。我们采用Cox模型与风险评分系统，在两项公开肝细胞癌队列（GSE76427与癌症基因组图谱（The Cancer Genome Atlas, TCGA））以及华山医院的一项队列中完成验证，总样本量共计641例。多因素Cox回归分析进一步证实，该风险评分可作为肝细胞癌复发预测的独立预后因子。此外，我们在两项验证队列的肝细胞癌组织中发现，ENY2、GPAA1及NDUFA4L2的表达水平显著上调；且ENY2在恶性肿瘤细胞中的表达量显著高于其余4个基因，提示ENY2可能在恶性细胞中发挥关键调控作用。细胞系实验结果显示，ENY2可促进细胞周期进程、细胞增殖、迁移与侵袭。对ENY2关联基因的功能分析表明，ENY2可能参与端粒维持这一癌症核心特征之一的调控过程。综上，本研究数据表明，ENY2或通过激活端粒维持通路调控肝细胞癌的恶性表型。