目的：筛选与肝细胞癌（hepatocellular carcinoma，HCC）预后相关的增强子RNA（enhancer RNA，eRNA）及其对应的 靶基因，并探究其调控作用及功能。方法：通过UCSC数据库下载HCC的转录本数据和临床病例数据，提取eRNA及其预测的 相应靶基因表达矩阵，用Kaplan-Meier和Spearman相关性分析方法筛选HCC预后相关的eRNA和靶基因，并采用最小化绝对收 缩和选择算子回归分析及多因素 Cox 多元回归分析构建 HCC 预后风险评分模型。设计合成针对筛选所获关键 eRNA AP003469.2的小干扰RNA并转染肝癌 SMMC-7721 细胞，采用 RT-PCR方法验证敲减效率，qPCR 法检测 AP003469.2 靶基因 YWHAZ的表达水平，CCK-8法检测转染后SMMC-7721细胞的增殖情况。结果：筛选获得4个与预后相关基因，包括两个eRNA （AP003469.2 和 SPRY4-AS1）和 两 个 靶 基 因（PPARGC1A 和 SLC2A1）（P<0.05），其中 PPARGC1A 高表达提示预后较好， AP003469.2、SPRY4-AS1和SLC2A1基因高表达则提示预后较差。基于4个基因构建的预后风险评分模型，第1、3和5年的AUC 分别为0.730、0.699和0.679，提示模型具备良好的预测效能。在敲减AP003469.2后，SMMC-7721细胞中YWHAZ的表达无明显 改变，且对细胞的增殖无影响。结论：基于生物信息学分析共筛选出4个关键基因，其中eRNA AP003469.2是HCC预后预测效 能较高的标志物，其无促癌功能且对YWHAZ基因无调控作用。

Objective: To screen the enhancer RNAs (eRNAs) and their corresponding target genes related to the prognosis hepatocellular carcinoma (HCC), and to explore their regulatory effect and function. Methods: The transcript data and clinical data of HCC were downloaded from UCSC database to extract the expression matrix of eRNAs and their predicted corresponding target genes. Kaplan-Meier and Spearman correlation analysis were used to screen HCC prognosis-related eRNAs and the target genes, and the least absolute shrinkage and selection operator (LASSO) regression analysis and multivariate Cox multiple regression analysis were utilized to construct a HCC prognostic risk scoring system. A short interfering RNA (siRNA) targeting eRNA AP003469.2 was designed and synthesized to transfect liver cancer SMMC-7721 cells. Then, RT-PCR was used to verify the knockdown efficiency, qPCR was used to detect the expression of the target gene YWHAZ, and CCK-8 was adopted to examine the proliferation of SMMC-7721 cells after transfection. Results: Four prognosis-related genes were screened, including two eRNAs (AP003469.2 and SPRY4-AS1) and two target genes (PPARGC1A and SLC2A1) (P<0.05). Among them, the high expression of PPARGC1A indicated a better prognosis, while the high expression of AP003469.2, SPRY4-AS1 and SLC2A1 genes indicated a poor prognosis. The prognostic risk scoring system constructed based on the 4 genes exhibited AUC of 0.730, 0.699 and 0.679 at the 1st, 3rd and the 5th year respectively, indicating good predictive performance of the system. After AP003469.2 knockdown in SMMC-7721 cells, the expression of its predicted target gene YWHAZ did not change significantly, and it had no significant effect on the proliferation of SMMC-7721 cells. Conclusion: Among the 4 key genes were screened out based on bioinformatics analysis, eRNA AP003469.2 is a biomarker with high prognostic performance for HCC; however, it has no cancer-promoting function or regulatory effect on YWHAZ.

天津市自然科学基金资助项目（No. 20JCYBJC01310）；天津市重大疾病防治科技重大专项（No. 19ZXDBSY00010）