摘要
目的 筛选结肠癌差异表达的 lncRNA和 miRNA,探讨其与结肠癌预后的关系及相关生物学功能.方法从TCGA数据库收集结肠癌患者的 lncRNA和 miRNA 表达谱数据及临床资料,利用 R 软件筛选差异表达的 ln-cRNA和 miRNA;单因素Cox回归分析与预后显著相关的差异表达 lncRNA和 miRNA;miRNet数据库构建预后相关的lncRNA-miRNA网络,利用mirPath v.3 软件对其进行功能富集分析;R软件构建预后风险模型并对其进行验证.采用CCK-8 实验、划痕实验、流式细胞术及 qPCR评估敲减 TSPEAR-AS2 对人结肠癌 HCT116 细胞增殖、迁移、凋亡及下游 miRNA表达的影响.结果 筛选出结肠癌组织中 1823 个差异表达的 lncRNA和 524 个差异表达的 miRNA;发现 158 个lncRNA和 31 个 miRNA与结肠癌的预后相关;构建由 8 个lncRNA和 14 个 miRNA 组成的 lncRNA-miRNA网络;GO 和KEGG富集分析显示,lncRNA-miRNA网络主要参与肿瘤相关的生物学功能及通路;鉴定出 11 个能有效评价结肠癌预后特征的关键基因,由这 11 个关键基因构建的预后风险模型中高风险组生存时间显著短于低风险组(P＜0.001),预测 1、3 和 5 年生存的AUC分别为 0.70、0.74 和 0.71.敲减TSPEAR-AS2可抑制 HCT116 细胞增殖、迁移、促进细胞凋亡,并下调 hsa-mir-4731-5p和 hsa-mir-342-3p 的表达.结论 成功筛选结肠癌相关 lncRNA-miRNA网络,并由此构建的预后风险模型具有良好的预测效能.TSPEAR-AS2 参与调控结肠癌细胞的增殖、迁移、凋亡以及下游 miRNA表达.
Abstract
Objective To screen differentially expressed lncRNA and miRNA in colon cancer,and to explore their association with colon cancer prognosis and related biological functions.Meth-ods The lncRNA and miRNA expression profile data and clinical information of colon cancer pa-tients were collected from TCGA database,and R software was used to screen differentially ex-pressed lncRNAs and miRNAs;Single-factor Cox regression analysis was used to identify differ-entially expressed lncRNA and miRNA that were significantly associated with prognosis;the miRNet database was used to construct a prognostic-related lncRNA-miRNA network;mirPath v.3 software was used for functional enrichment analysis;R software was used to build a prog-nostic risk model and validate it.CCK-8 experiments,scratch experiments,flow cytometry,and qPCR were used to evaluate the effects of knocking down TSPEAR-AS2 on the proliferation,mi-gration,apoptosis and downstream miRNA expression of human colon cancer HCT116 cells.Re-sults 1823 differentially expressed lncRNAs and 524 differentially expressed miRNAs were iden-tified in colon cancer tissues;158 lncRNAs and 31 miRNAs were found to be associated with co-lon cancer prognosis;an lncRNA-miRNA network composed of 8 lncRNAs and 14 miRNAs was constructed;GO and KEGG enrichment analyses showed that the lncRNA-miRNA network was mainly involved in tumor-related biological functions and pathways;11 key genes that could effec-tively evaluate colon cancer prognostic characteristics were identified,the survival time of the high-risk group was significantly shorter than that of the low-risk group in the prognostic risk model constructed from these 11 key genes(P＜0.001),and the AUCs for predicting the survival of 1,3,and 5 years were 0.70,0.74,and 0.71,respectively.Knocking down TSPEAR-AS2 inhibited the proliferation,migration,and promoted apoptosis of HCT116 cells,and down-regulated the ex-pression of hsa-mir-4731-5p and hsa-mir-342-3p.Conclusion successful screening of colon cancer-associated lncRNA-miRNA network and the prognostic risk model constructed therefrom can has good predictive efficacy.TSPEAR-AS2 could be involved in regulating the proliferation,migra-tion,apoptosis,and downstream miRNA expression of colon cancer cells.
基金项目
江西省自然科学基金(20202BABL206802)
维普
万方数据