Department of Medical Laboratory, First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450052, P.R. China.
Department of Medical Laboratory, The Second Affiliated Hospital of Henan University of Traditional Chinese Medicine, Zhengzhou, Henan 450000, P.R. China.
Oncol Rep. 2020 Sep;44(3):987-1002. doi: 10.3892/or.2020.7658. Epub 2020 Jun 23.
Esophageal squamous cell carcinoma (ESCC), the main subtype of esophageal cancer (EC), is a common lethal type of cancer with a high mortality rate. The aim of the present study was to select key relevant genes and identify potential mechanisms involved in the development of ESCC based on bioinformatics analysis. Minichromosome maintenance 6 complex component (MCM6) has been identified to be upregulated in multiple malignancies; however, its contributions to ESCC remain unclear. For the purposes of the present study, four datasets were downloaded from the Gene Expression Omnibus (GSE63941, GSE26886, GSE17351 and GSE77861), and the intersection of the differentially expressed genes was obtained using a Venn diagram. The protein‑protein interaction was then constructed, and the modules were verified by Cytoscape, in which the key genes have a high connectivity degree with other genes. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway were subsequently filtered out to analyze the development of ESCC. MCM6, an upregulated gene, was selected and connected with most of the other genes, for further research validation. The expression levels of MCM6 were then assessed using the Oncomine, GEPIA and UALCAN databases and validated in both ESCC tissues samples and cell lines by immunohistochemistry and RT‑qPCR. Cell counting kit‑8 (CCK‑8), flow cytometry, wound healing and Transwell assays were used to determine the proliferation, apoptosis, cell cycle, migration and invasion of ESCC cells. A total of 24 genes were identified by a series of bioinformatics analyses and the results revealed that the genes were associated with DNA replication and cell cycle. Experimental validation revealed that MCM6 expression was significantly elevated in both ESCC tissues and cell lines. The results were consistent with those of bioinformatics analysis. Furthermore, the knockdown of MCM6 inhibited cell proliferation, migration and invasion and promoted cell apoptosis, and made cells arrested in S stage. In summary, the findings of bioinformatics analysis provided a novel hypothesis for ESCC progression. In particular, the aberrantly elevated expression of MCM6 is a potential biomarker for ESCC diagnosis and treatment.
食管鳞状细胞癌 (ESCC) 是食管癌 (EC) 的主要亚型,是一种常见的致死性癌症,死亡率很高。本研究旨在通过生物信息学分析,选择关键的相关基因,并鉴定 ESCC 发生发展中的潜在机制。微小染色体维持 6 复合物成分 (MCM6) 在多种恶性肿瘤中被鉴定为上调;然而,其对 ESCC 的贡献尚不清楚。在本研究中,从基因表达综合 (GEO) 数据库中下载了四个数据集 (GSE63941、GSE26886、GSE17351 和 GSE77861),并使用 Venn 图获取差异表达基因的交集。然后构建蛋白质-蛋白质相互作用,使用 Cytoscape 验证模块,其中关键基因与其他基因具有较高的连接度。随后进行基因本体论和京都基因与基因组百科全书通路筛选,以分析 ESCC 的发生发展。选择上调基因 MCM6,并与大多数其他基因连接,进行进一步的研究验证。使用 Oncomine、GEPIA 和 UALCAN 数据库评估 MCM6 的表达水平,并通过免疫组化和 RT-qPCR 在 ESCC 组织样本和细胞系中进行验证。细胞计数试剂盒-8 (CCK-8)、流式细胞术、划痕愈合和 Transwell 实验用于测定 ESCC 细胞的增殖、凋亡、细胞周期、迁移和侵袭。通过一系列生物信息学分析鉴定出 24 个基因,结果表明这些基因与 DNA 复制和细胞周期有关。实验验证表明,MCM6 在 ESCC 组织和细胞系中的表达均显著升高。结果与生物信息学分析一致。此外,MCM6 的敲低抑制了细胞增殖、迁移和侵袭,促进了细胞凋亡,并使细胞停滞在 S 期。总之,生物信息学分析的结果为 ESCC 进展提供了一个新的假说。特别是,MCM6 的异常高表达可能成为 ESCC 诊断和治疗的潜在生物标志物。