背景:NME1的高表达与肝细胞癌(HCC)的进展和不良预后相关。然而,关于NME1和microRNAs(miRNA)在HCC进展中的相关性的报道很少。
目的:探索调节HCC中NME1表达的miRNA。
方法:来自癌症基因组图谱(TCGA)的数据,人蛋白质图谱(HPA),TargetScan,starBase,和mirDIP用于分析NME1在HCC组织中的表达模式,NME1水平与HCC进展或患者预后的关系,靶向NME1的miRNA,以及可能受NME1调控的生物过程。使用双荧光素酶报告基因测定评估miRNA对NME1的调节,定量逆转录聚合酶链反应(qRT-PCR)和蛋白质印迹。用碘化丙啶(PI)染色和EdU法检测细胞周期和细胞增殖,分别。
结果:NME1在HCC中高表达与HCC进展和预后相关。miR-139-5p和miR-335-5p在肝癌标本中呈弱表达,与NME1呈负相关。miR-139-5p在HCC患者中的下调导致更差的总生存期(OS)和无病间隔(DFI);然而,miR-335-5p水平与HCC患者的OS和DFI无显著相关性。体外实验证实,与L-02相比,HCC细胞系中miR-139-5p的水平较低,NME1表达较高。此外,miR-139-5p负调控NME1在HCC细胞系中的表达。NME1可能调节细胞周期,DNA复制,氧化磷酸化,和磷酸戊糖途径。miR-139-5p通过负调控NME1表达抑制细胞增殖。
结论:NME1在HCC中的上调提示预后不良。NME1受miR-139-5p负调控以抑制细胞增殖。
BACKGROUND: High expression of NME1 is associated with hepatocellular carcinoma (HCC) progression and poor prognosis. However, there are few reports on the association between NME1 and microRNAs (miRNAs) in HCC progression.
OBJECTIVE: To explore miRNAs that regulate
NME1 expression in HCC.
METHODS: Data from the Cancer Genome Atlas (TCGA), Human Protein Atlas (HPA), TargetScan, starBase, and mirDIP were used to analyze the expression pattern of NME1 in HCC tissues, the relationship between NME1 level and the progression of HCC or patient prognosis, miRNAs targeting
NME1, and the biological processes that may be regulated by
NME1. The regulation of miRNAs to NME1 was assessed using the dual-luciferase reporter assay, quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blotting. The cell cycle and cell proliferation were detected using propidium iodide (PI) staining and EdU assay, respectively.
RESULTS: Highly expressed NME1 in HCC was associated with HCC progression and prognosis. The miR-139-5p and miR-335-5p were weakly expressed in HCC samples and negatively correlated with NME1. The downregulation of miR-139-5p in HCC patients resulted in worse overall survival (OS) and disease-free interval (DFI); however, the level of miR-335-5p was not significantly correlated with OS and DFI in patients with HCC. In vitro experiments verified that the level of miR-139-5p was lower and NME1 expression was higher in HCC cell lines compared to L-02. Moreover, miR-139-5p negatively regulates the expression of
NME1 in HCC cell lines. The NME1 may regulate cell cycle, DNA replication, oxidative phosphorylation, and the pentose phosphate pathway. The miR-139-5p inhibited cell proliferation by negatively regulating NME1 expression.
CONCLUSIONS: The upregulation of
NME1 in HCC indicates a poor prognosis. The
NME1 is negatively regulated by miR-139-5p to inhibit cell proliferation.