摘要
目的:分析EGLN3在肺腺癌(lung adenocarcinoma, LUAD)中的作用。方法:(1)通过癌症基因组图谱(the cancer genome atlas, TCGA)数据库下载535例肺腺癌样本和59例正常样本，分析EGLN3在肺腺癌和正常肺组织中的表达差异；(2)使用R语言分析EGLN3的表达水平及其在肺腺癌中的临床意义；(3)通过Kaplan-Meier方法、单因素和多因素COX回归分析和生存预测列线图确定EGLN3的表达水平与肺腺癌预后的关系；(4)通过基因集富集分析(GSEA)筛选EGLN3相关的生物学途径；(5)细胞实验：通过蛋白质印迹法(WB)验证EGLN3在肺腺癌细胞中是否存在差异表达；对EGLN3基因进行过表达和干扰表达，采用CCK-8增殖实验、划痕实验和Transwell侵袭实验，明确表达水平对肺腺癌细胞增殖、迁移和侵袭的影响。结果:(1)EGLN3在肺腺癌组织中表达水平高于正常肺组织细胞；(2)EGLN3表达水平与肺腺癌病理分期相关，病理分期越高，对应EGLN3表达水平越高；(3)EGLN3表达水平与肺腺癌总生存期(OS)独立预后相关，EGLN3高表达肺腺癌较EGLN3低表达肺腺癌OS、疾病特异性生存期(DSS)和无进展间隔(PFI)差；(4)基因集富集分析预测EGLN3可能参与的信号通路包括激活MYC基因、上皮细胞-间充质转化(EMT)、DNA复制、细胞周期、mTORC1通路、G2/M检查点、E2F转录过程、P53信号通路等；(5)细胞实验：EGLN3在肺腺癌细胞中表达水平高于人支气管上皮样细胞；CCK-8增殖实验、划痕实验和Transwell侵袭实验显示，EGLN3过表达促进肺腺癌细胞增殖、侵袭和迁移，EGLN3干扰表达抑制肺腺癌细胞增殖、侵袭和迁移。结论:EGLN3在肺腺癌细胞中存在高表达，提示肺腺癌预后不良，可作为肺腺癌预后的预测因素。EGLN3表达可促进肺腺癌细胞增殖、侵袭和迁移。
Abstract
Objective:EGLN3 is differentially expressed in many kinds of tumors. However, the role of EGLN3 in lung adenocarcinoma (LUAD) is not clear. The purpose of this study is to explore the role of EGLN3 in lung adenocarcinoma.Methods:(1)535 LUAD samples and 59 normal samples were downloaded from Cancer Genome Map (TCGA) database to analyze the difference of EGLN3 expression between LUAD and normal lung tissues. (2)The expression level of EGLN3 and its clinical significance in LUAD were analyzed by R language. (3)The relationship between the expression of EGLN3 and the prognosis of LUAD was determined by Kaplan-Meier method, univariate and multivariate COX regression analysis and survival prediction diagram. (4)The biological pathways related to EGLN3 were screened by gene sets enrichment analysis (GSEA). (5)Cell experiment: Western blotting (WB) was used to verify the differential expression of EGLN3 in lung adenocarcinoma cells, and then EGLN3 was overexpressed and interfered, and CCK-8 proliferation test, scratch test and Transwell invasion test were carried out to determine the effect of its expression level on the proliferation, migration and invasion of lung adenocarcinoma cells.Results:The expression level of EGLN3 in LUAD tissue was significantly higher than that in normal lung tissue. (2)The expression level of EGLN3 is related to the pathological stage of lung adenocarcinoma. The higher the pathological stage, the higher the expression level of EGLN3. (3)The expression of EGLN3 is an independent prognostic factor related to the overall survival (OS) of lung adenocarcinoma. The patients with high expression of EGLN3 had worse OS, disease specific survival (DSS) and progress free interval (PFI) than those with low expression of EGLN3. (4)Gene sets enrichment analysis predicted the possible signal pathways involved in EGLN3, including activation of MYC gene, epithelial mesenchymal transformation (EMT), DNA replication, cell cycle, mTORC1 pathway, G2/M checkpoint, E2F transcription process, p53 signal pathway, etc. (5) Cell experiment: The expression level of EGLN3 in LUAD cells was significantly higher than that in human bronchial epithelioid cells. CCK-8 proliferation test, scratch test and Transwell invasion test: the overexpression of EGLN3 promoted the proliferation, invasion and migration of lung adenocarcinoma cells, while the interference expression of EGLN3 inhibited the proliferation, invasion and migration of lung adenocarcinoma cells.Conclusions:EGLN3 is highly expressed in lung adenocarcinoma cells, and its high expression indicates a poor prognosis of lung adenocarcinoma, which can be used as an independent factor to predict the prognosis of lung adenocarcinoma. The expression of EGLN3 can promote the proliferation, invasion and migration of lung adenocarcinoma cells.
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