Department of Gastroenterology and Hepatology, Zhongshan Hospital of Fudan University, Shanghai 200032, P.R. China.
Division of Gastroenterology, University of Toronto and University Health Network, Toronto, ON M5G 2C4, Canada.
Int J Mol Med. 2019 Nov;44(5):1844-1854. doi: 10.3892/ijmm.2019.4323. Epub 2019 Aug 23.
Long non‑coding RNAs (lncRNAs) have been shown to contribute to progression and prognosis of hepatocellular carcinoma (HCC). However, expression profiling and interaction of lncRNAs with messenger RNAs (mRNAs) and microRNAs (miRNAs) remain largely unknown in HCC. The expression profiling of lncRNAs, mRNA and miRNAs was obtained using microarray. The Gene Ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes pathway analysis were used to characterize potential functions of differentially expressed mRNAs. Cytoscape was applied to construct an lncRNA‑miRNA‑mRNA co‑expression network and candidate lncRNAs were validated via quantitative PCR in 30 pairs of HCC and adjacent tumor‑free tissues. In this study, 1,056 upregulated and 1,288 downregulated lncRNAs were identified, while 2,687 mRNAs and 6 miRNAs were aberrantly expressed in HCC compared with adjacent tumor‑free tissues. Potential functions of differentially expressed mRNAs were demonstrated to significantly participate in modulating critical genes in the cell cycle, such as cyclin E1 and cyclin B2. After screening, 95 lncRNAs, 5 miRNAs and 36 mRNAs were recruited for construction of lncRNA‑mRNA‑miRNA co‑expression network in the cell cycle pathway. Subsequently, the top 5 lncRNAs that potentially modulate critical genes in the cell cycle were selected as the candidates for further verification. Kaplan‑Meier curves using the Cancer Genome Atlas database showed that 13 targeted mRNAs were associated with overall survival of HCC patients. Finally, three lncRNAs, including ENST00000522221, lnc‑HACE1‑6:1 and lnc‑ICOSLG‑11:1, are significantly upregulated in HCC tissues compared with adjacent tumor‑free tissues. These findings suggest that lncRNAs play essential roles in the pathogenesis of HCC via regulating coding genes and miRNAs, and may be important targets for diagnosis and treatment of this disease.
长链非编码 RNA（lncRNA）已被证明与肝细胞癌（HCC）的进展和预后有关。然而，lncRNA 与信使 RNA（mRNA）和 microRNA（miRNA）的表达谱和相互作用在 HCC 中仍知之甚少。使用微阵列获得 lncRNA、mRNA 和 miRNA 的表达谱。基因本体论（GO）富集和京都基因与基因组百科全书通路分析用于表征差异表达 mRNA 的潜在功能。Cytoscape 用于构建 lncRNA-miRNA-mRNA 共表达网络，并通过定量 PCR 在 30 对 HCC 和相邻无肿瘤组织中验证候选 lncRNA。在这项研究中，与相邻无肿瘤组织相比，HCC 中鉴定出 1056 个上调和 1288 个下调的 lncRNA，同时有 2687 个 mRNA 和 6 个 miRNA 异常表达。差异表达 mRNA 的潜在功能表明其显著参与调节细胞周期中的关键基因，如周期蛋白 E1 和周期蛋白 B2。筛选后，在细胞周期通路中构建 lncRNA-mRNA-miRNA 共表达网络，共招募 95 个 lncRNA、5 个 miRNA 和 36 个 mRNA。随后，选择 5 个可能调节细胞周期中关键基因的 top5 lncRNA 作为进一步验证的候选物。使用癌症基因组图谱数据库的 Kaplan-Meier 曲线显示，13 个靶向 mRNA 与 HCC 患者的总生存率相关。最后，与相邻无肿瘤组织相比，在 HCC 组织中，三个 lncRNA，包括 ENST00000522221、lnc-HACE1-6:1 和 lnc-ICOSLG-11:1，显著上调。这些发现表明，lncRNA 通过调节编码基因和 miRNA 在 HCC 的发病机制中发挥重要作用，可能是该疾病诊断和治疗的重要靶点。
