肝细胞癌患者预后相关增强子RNA和对应靶基因的筛选及其预后预测意义  

Screening of prognosis-related enhancer RNAs and their corresponding target genes in patients with hepatocellular carcinoma and its prognostic significance

作　　者：白易 吴昊[2] 张雅敏[1] BAI Yi;WU Hao;ZHANG Yamin(Department of Hepatobiliary Surgery,the First Central Hospital of Tianjin City,Tianjin 300192,China;Department of Hepatobiliary Surgery,the First Center Clinical College of Tianjin Medical University,Tianjin 300192,China)

机构地区：[1]天津市第一中心医院肝胆外科,天津300192 [2]天津医科大学第一中心临床学院肝胆外科,天津300192

出　　处：《中国肿瘤生物治疗杂志》2021年第9期914-919,共6页Chinese Journal of Cancer Biotherapy

基　　金：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)。

摘　　要：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.

关 键 词：肝细胞癌 SMMC-7721细胞 生物信息学 预后 增强子RNA 靶基因 

分 类 号：R735.7[医药卫生—肿瘤] R730.7[医药卫生—临床医学]