Identification of prognostic markers of lung cancer through bioinformatics analysis and in vitro experiments
Identification of prognostic markers of lung cancer through bioinformatics analysis and in vitro experiments
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通过生物信息学分析和体外实验鉴定肺癌预后标志物
DOI：
10.3892/ijo.2019.4926
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发表时间：
2020-01-01
影响因子：
5.2
通讯作者：
Qi, Guangzi
中科院分区：
文献类型：
--
作者：
Ling, Bo;Liao, Xianjiu;Qi, Guangzi
肺癌是世界上最常见的癌症类型之一。了解肺癌发生和发展的分子机制可能有助于早期诊断、治疗和预后。本研究的目的是研究肺癌的发病机制，并确定潜在的新的生物标志物。肺癌患者的基因表达数据集从Gene Expression Omnibus获得。通过加权基因共表达网络分析筛选与肺癌关系最密切的基因（核心基因）。进一步利用体外细胞实验来验证核心基因对肺癌细胞增殖、细胞与基质之间的粘附以及相关代谢途径的影响。基于WGCNA筛选，确定了与肺癌密切相关的2个基因模块和5个核心基因，包括来自绿松石模块的免疫球蛋白超家族成员10（IGSF 10）和来自棕色模块的核糖核苷酸还原酶调节亚基M2、胞质分裂蛋白调节因子1、驱动蛋白家族成员（KIF）14和KIF 2C。生存分析和差异基因表达分析显示IGSF 10在健康对照组和肺癌患者中的表达水平存在显著差异。在肺癌患者中，IGSF 10表达降低，肺癌患者的总生存时间明显缩短。MTT和集落形成实验表明IGSF 10基因敲除后肺癌细胞增殖明显增加，Transwell实验和粘附实验进一步表明IGSF 10基因敲除后细胞与基质的粘附明显增加。基因集富集分析显示IGSF 10的表达水平与整合素-β1/黏着斑激酶（FAK）通路的激活密切相关。Western blotting结果显示，IGSF 10基因敲除后，整合素β1/FAK通路激活，整合素β1、磷酸化（p）-FAK和p-AKT蛋白表达水平显著上调。IGSF 10基因敲除后整合素β1/FAK通路的激活影响肺癌细胞的增殖和粘附。IGSF 10可作为肺癌的一个潜在的预后指标。
Lung cancer is one of the most common types of cancer worldwide. Understanding the molecular mechanisms underlying the development and progression of lung cancer may improve early diagnosis, treatment and prognosis. The aim of the present study was to examine the pathogenesis of lung cancer and to identify potentially novel biomarkers. Gene expression datasets of patients with lung cancer were obtained from the Gene Expression Omnibus. Genes which were most closely associated with lung cancer (core genes) were screened by weighted gene co-expression network analysis. In vitro cell based experiments were further utilized to verify the effects of the core genes on the proliferation of lung cancer cells, adhesion between cells and the matrix, and the associated metabolic pathways. Based on WGCNA screening, two gene modules and five core genes closely associated with lung cancer, including immunoglobulin superfamily member 10 (IGSF10) from the turquoise module, and ribonucleotide reductase regulatory subunit M2, protein regulator of cytokinesis 1, kinesin family member (KIF)14 and KIF2C from the brown module were identified as relevant. Survival analysis and differential gene expression analysis showed that there were significant differences in IGSF10 expression levels between the healthy controls and patients with lung cancer. In patients with lung cancer, IGSF10 expression was decreased, and the overall survival time of patients with lung cancer was significantly shortened. An MTT and colony formation assay showed that IGSF10-knockout significantly increased proliferation of lung cancer cells, and Transwell assays and adhesion experiments further suggested that the adhesion between cells and the matrix was significantly increased in IGSF10-knockout cells. Gene Set Enrichment Analysis showed that the expression level of IGSF10 was significantly associated with the activation of the integrin-β1/focal adhesion kinase (FAK) pathway. Western blotting revealed that knockout of IGSF10 resulted in the activation of the integrin-β1/FAK pathway, as the protein expression levels of integrin-β1, phosphorylated (p)-FAK and p-AKT were significantly upregulated. Activation of the integrin-β1/FAK pathway, following knockout of IGSF10, affected the proliferation and adhesion of lung cancer cells. Therefore, IGSF10 my serve as a potential prognostic marker of lung cancer.