目的:利用癌症基因组图谱(The Cancer Genome Atlas,TCGA)数据库筛选肝细胞癌(hepatocellular carcinoma,HCC)组织中与失巢凋亡相关的miRNAs(anoikis-related miRNAs,armiRNAs),基于筛选出来的miRNAs构建HCC预后列线图模型,探讨armiRNAs与HCC预后的相关性,为HCC的治疗提供新的靶点.方法:从TCGA数据库下载RNA转录数据,筛选在HCC和癌旁组织中差异表达的armiRNAs,采用单因素Cox回归和LASSO回归分析鉴定具有预后意义的armiRNAs,通过受试者工作特征曲线(ROC曲线)评价其可靠性,并构建预后列线图模型.利用该模型,使用京都基因和基因组百科全书(KEGG)进行基因富集分析及药物敏感性研究.通过RT-qPCR对人正常肝细胞系LO2和三种人肝癌细胞系中差异表达的armiR-NAs 进行验证.结果:本研究经过筛选得到7个armiRNAs,根据风险评分将肝癌样本分为高、低风险组,Kaplan-Meier生存曲线显示两组之间存在显著差异,死亡人数随着风险评分的增加而增加.预测1年、2年、3年生存率的ROC曲线的AUC值分别为0.794、0.789、0.781,表明该模型具有较高的区分度.针对不同风险组对抗肿瘤药物敏感性的研究结果表明,高危组对多种抗肿瘤药物敏感性较高.实验验证显示,与LO2细胞相比,三种人肝癌细胞系中hsa-miR-4661、hsa-miR-1180表达水平显著增高,hsa-miR-139、hsa-miR-101-2、hsa-miR-326、hsa-miR-29c 表达水平显著下降,而 hsa-miR-509-3-5p 在HepG2细胞中表达显著降低,在SMMC-7721、MHCC97细胞中表达显著升高.结论:本研究共筛选出7个armiRNAs,基于这些基因构建的模型能够较好的预测HCC患者的生存率及患者对药物的敏感性,这些结果可为探索肝癌预后标志物和临床个体化治疗提供重要参考.
Objective:Use The Cancer Genome Atlas(TCGA)database to screen anoikis-related miRNAs(armiRNAs)in hepatocellular carcinoma(HCC)tissues and construct a prognostic model based on the selected miRNAs.To explored the relevance of armiRNAs and the prognosis of HCC,hope to provide new targets for liver canc-er therapy.Methods:RNA transcription data were downloaded from the TCGA.The differentially expressed armiRNAs were screened in HCC and paraneoplastic tissues.Univariate Cox regression and LASSO regression analyses were used to identify prognostically significant armiRNAs.The reliability was assessed by ROC curves,then a prognostic nomo-gram model was constructed.Using this model,gene enrichment analysis and drug susceptibility studies were per-formed using the Kyoto Gene and Genome Encyclopedia(KEGG).Validation of differentially expressed armiRNAs in human normal hepatocyte cell line LO2 and three human hepatocellular carcinoma cell lines by RT-qPCR.Results:This study screened seven armiRNAs,then classified the liver cancer samples into high-risk and low-risk groups according to risk scores.Kaplan-Meier survival curves showed significant differences between the two groups.Mortal-ity increased with risk scores.The AUC of the ROC curves predicting 1-,2-and 3-year survival were 0.794,0.789,and 0.781,indicating a higher degree of discrimination in the model.High-risk groups had higher suscepti-bility to multiple anti-tumour drugs.Compared with LO2 cells,the expression of hsa-miR-4661,hsa-miR-1180 was significantly increased in three kinds of human HCC cell lines,the expression of hsa-miR-139,hsa-miR-101-2,hsa-miR-326,and hsa-miR-29c was significantly decreased,whereas hsa-miR-509-3-5p was sig-nificantly reduced in HepG2 cells and significantly increased in SMMC-7721 and MHCC97 cells.Conclusion:This study identified seven armiRNAs.Models which based on these genes are good predictors of survival and drug sensitiv-ity in patients with HCC.These results may provide some guidance for the exploration of prognostic markers and clini-cal individualized treatment of HCC.