Institute of Pathology, Faculty of Medicine, University of Ljubljana, Korytkova 2, 1000 Ljubljana, Slovenia.
Department of Otorhinolaryngology and Cervicofacial Surgery, University Medical Centre Ljubljana, Zaloška 2 and Faculty of Medicine, University of Ljubljana, Korytkova 2, 1000 Ljubljana, Slovenia.
Biomed Res Int. 2020 Jul 17;2020:8573793. doi: 10.1155/2020/8573793. eCollection 2020.
Results of previous studies suggest that NANOG may be an important prognostic biomarker in oral squamous cell carcinoma (OSCC), but there are contradictory results regarding NANOG expression patterns on mRNA and protein levels, and the mechanisms of its regulation are poorly understood. Our aim was to analyze the expression and diagnostic significance of NANOG in OSCC, and the possible mechanisms of its regulation, i.e., protein regulators on mRNA level (, , , , and ), methylation status, copy number variation, and regulatory miRNAs, , , , , , and .
Our study included 120 patients with OSCC. Expression of NANOG protein and mRNA was analyzed using immunohistochemistry and qPCR. Expression of regulatory factors, miRNAs, and copy number variation was performed using qPCR. Methylation status of promoter was determined using PCR and Sanger sequencing.
We detected upregulation of and and downregulation of and mRNA in OSCC with lymph node metastases compared to OSCC without lymph node metastases. We observed a strong positive correlation between mRNAs of and those of its protein regulators , , , , and . The expression of was in positive correlation with the expression of . There was also a correlation between T status of OSCC and the expression of and and a correlation of with the N status of T2 OSCC. NANOG promoter methylation and copy number variation were only observed in a small proportion of samples.
Our findings confirm the diagnostic significance of NANOG in OSCC and provide information on NANOG expression patterns on both mRNA and protein levels. They also suggest that protein regulators and microRNAs might play a crucial role, whereas methylation of its promoter and copy number variation probably have a minor role in the regulation of NANOG expression in OSCC.
先前的研究结果表明，NANOG 可能是口腔鳞状细胞癌（OSCC）的一个重要预后生物标志物，但关于 NANOG 在 mRNA 和蛋白水平上的表达模式存在矛盾的结果，其调控机制也知之甚少。我们的目的是分析 NANOG 在 OSCC 中的表达和诊断意义，以及其可能的调控机制，即在 mRNA 水平上的蛋白调控因子（、、、、和）、甲基化状态、拷贝数变异和调控 miRNA（、、、、、、和）。
我们的研究包括 120 例 OSCC 患者。使用免疫组织化学和 qPCR 分析 NANOG 蛋白和 mRNA 的表达。使用 qPCR 分析调控因子、miRNA 和拷贝数变异的表达。使用 PCR 和 Sanger 测序法确定启动子的甲基化状态。
我们检测到具有淋巴结转移的 OSCC 中上调和 ，下调和 。我们观察到和其蛋白调控因子、、、和之间的 mRNA 存在强烈的正相关。的表达与的表达呈正相关。OSCC 的 T 分期与和的表达以及与 T2 OSCC 的 N 分期之间也存在相关性。NANOG 启动子甲基化和拷贝数变异仅在一小部分样本中观察到。
我们的发现证实了 NANOG 在 OSCC 中的诊断意义，并提供了关于 NANOG 在 mRNA 和蛋白水平上表达模式的信息。它们还表明，蛋白调控因子和 microRNAs 可能发挥关键作用，而其启动子的甲基化和拷贝数变异可能在 OSCC 中 NANOG 表达的调控中起次要作用。
