Early diagnosis and prognosis of hepatocellular carcinoma based on a ceRNA array
Early diagnosis and prognosis of hepatocellular carcinoma based on a ceRNA array
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DOI：
10.1515/oncologie-2023-0088
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发表时间：
2023-03
期刊：
影响因子：
0.9
通讯作者：
Lixin Wang;A. Kong;Hui Dong
中科院分区：
文献类型：
--
作者：
Lixin Wang;A. Kong;Hui Dong
摘要 目的 肝细胞癌（HCC）是癌症相关死亡最常见的原因之一，其发病率高、早期诊断率低、预后差。探索竞争性内源性 RNA (ceRNA) 芯片对于 HCC 的早期诊断和预后至关重要。方法从TCGA数据库下载HCC中差异表达的lncRNA、miRNA和mRNA的原始基因表达谱。从miRcode和starBase中提取差异表达的lncRNA-miRNA和miRNA-mRNA相互作用对，构建ceRNA网络，并进行GO注释和KEGG通路分析。 Cox 回归和 Kaplan-Meier 生存分析筛选确定了与 HCC 生存相关的网络中的核心基因，以 miRNA 为中心，并使用 ceRNA 阵列进行筛选。采用qRT-PCR检测临床样本中关键基因的表达水平。双荧光素酶报告基因检测用于验证lncRNA、mRNA和miRNA之间的靶结合关系。 ROC曲线用于分析ceRNA阵列的诊断功效。结果 总共使用8个lncRNA、5个miRNA和21个mRNA构建ceRNA网络。功能富集分析显示，ceRNA网络中的mRNA主要富集于14条信号通路，尤其是癌症中的microRNA。生存分析显示lncRNA FOXD2-AS1和miRNA miR-9-5p与HCC的预后相关，以及mRNA之间的靶向结合关系。来自 TargetScan、starBase、miRDB 和 PicTar 数据库的 STMN1、COL15A1 和 CCNE2 以及 miR-9-5p 是可靠的。 qRT-PCR 显示 HCC 组织中 FOXD2-AS1、miR-9-5p、STMN1、COL15A1 和 CCNE2 的表达水平上调。双荧光素酶报告基因检测显示 FOXD2-AS1 和 STMN1 与 miR-9-5p 具有靶向结合关系，但与 COL15A1 或 CCNE2 没有靶向结合关系。候选ceRNA阵列（FOXD2-AS1/miR-9-5p/STMN1/COL15A1/CCNE2）的曲线下面积高于每个成员和ceRNA组合（FOXD2-AS1/miR-9-5p/STMN1）的曲线下面积。结论 FOXD2-AS1/miR-9-5p/STMN1/COL15A1/CCNE2形成的候选ceRNA芯片可作为HCC早期诊断和预后的生物标志物。
Abstract Objectives Hepatocellular carcinoma (HCC) is one of the most common causes of cancer-related deaths, due to high morbidity, a low early diagnosis rate, and poor prognosis. It is essential to explore competitive endogenous RNA (ceRNA) arrays for early diagnosis and prognosis of HCC. Methods The original gene expression profiles of differentially expressed lncRNA, miRNA and mRNA in HCC were downloaded from TCGA database. Differentially expressed lncRNA-miRNA and miRNA-mRNA interaction pairs were extracted from miRcode and starBase, a ceRNA network was constructed, and GO annotation and KEGG pathway analyses were performed. Cox regression and Kaplan-Meier survival analysis screening identified core genes in the network associated with HCC survival, centering on miRNA, which were screened using ceRNA arrays. qRT-PCR was used to detect the expression level of key genes in clinical samples. Dual luciferase reporter gene assays were used to verify the target binding relationship among lncRNA, mRNA, and miRNA. ROC curves were used to analyze the diagnostic efficacy of the ceRNA array. Results A total of 8 lncRNAs, 5 miRNAs, and 21 mRNAs were used to construct a ceRNA network. Functional enrichment analysis showed that mRNAs in the ceRNA network were mainly enriched in 14 signaling pathways, especially microRNAs in cancer. Survival analysis showed that lncRNA FOXD2-AS1 and miRNA miR-9-5p were related to the prognosis of HCC, and the targeted binding relationships between mRNAs. STMN1, COL15A1, and CCNE2 and miR-9-5p from the TargetScan, starBase, miRDB, and PicTar databases were reliable. qRT-PCR showed that expression levels of FOXD2-AS1, miR-9-5p, STMN1, COL15A1, and CCNE2 were upregulated in HCC tissues. Dual luciferase reporter assays showed that FOXD2-AS1 and STMN1 had a targeted binding relationship with miR-9-5p, but not with COL15A1 or CCNE2. The area under the curve of the candidate ceRNA array (FOXD2-AS1/miR-9-5p/STMN1/COL15A1/CCNE2) was higher than that of each member and ceRNA combination (FOXD2-AS1/miR-9-5p/STMN1). Conclusions The candidate ceRNA array formed by FOXD2-AS1/miR-9-5p/STMN1/COL15A1/CCNE2 could be a biomarker for early diagnosis and prognosis of HCC.