乙型肝炎病毒相关肝细胞癌中p53信号通路三个核心基因的鉴定与验证

Identification and validation of three core genes in p53 signaling pathway in hepatitis B virus-related hepatocellular carcinoma.

作者信息

Yu Mingxue, Xu Wenli, Jie Yusheng, Pang Jiahui, Huang Siqi, Cao Jing, Gong Jiao, Li Xinhua, Chong Yutian

机构信息

Department of Infectious Diseases and Key Laboratory of Liver Disease of Guangdong Province, Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, 510630, Guangdong Province, China.

Department of Laboratory Medicine, Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, 510630, Guangdong Province, China.

出版信息

World J Surg Oncol. 2021 Mar 8;19(1):66. doi: 10.1186/s12957-021-02174-w.

DOI:10.1186/s12957-021-02174-w

PMID:33685467

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7938465/

Abstract

BACKGROUND

Hepatocellular carcinoma (HCC) is a common cancer and the leading cause is persistent hepatitis B virus (HBV) infection. We aimed to identify some core genes and pathways for HBV-related HCC.

METHODS

Gene expression profiles of GSE62232, GSE121248, and GSE94660 were available from Gene Expression Omnibus (GEO). The GSE62232 and GSE121248 profiles were the analysis datasets and GSE94660 was the validation dataset. The GEO2R online tool and Venn diagram software were applied to analyze commonly differentially expressed genes between HBV-related HCC tissues and normal tissues. Then, functional enrichment analysis using Gene Ontology (GO) and the Kyoto Encyclopedia of Gene and Genome (KEGG) as well as the protein-protein interaction (PPI) network was conducted. The overall survival rates and the expression levels were detected by Kaplan-Meier plotter and Gene Expression Profiling Interactive Analysis (GEPIA). Next, gene set enrichment analysis (GSEA) was performed to verify the KEGG pathway analysis. Furthermore, quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) was performed to validate the levels of these three core genes in tumor tissues and adjacent non-tumor liver tissues from 12 HBV related HCC patients, HBV-associated liver cancer cell lines and normal liver cell lines, and HepG2 with p53 knockdown or deletion, respectively.

RESULTS

Fifteen highly expressed genes associated with significantly worse prognoses were selected and CCNB1, CDK1, and RRM2 in the p53 signaling pathway were identified as core genes. GSEA results showed that samples highly expressing three core genes were all enriched in the p53 signaling pathway in a validation dataset (P < 0.0001). The expression of these three core genes in tumor tissue samples was higher than that in relevant adjacent non-tumor liver tissues (P < 0.0001). Furthermore, we also found that the above genes were highly expressed in liver cancer cell lines compared with normal liver cells. In addition, we found that the expression of these three core genes in p53 knockdown or knockout HCC cell lines was lower than that in negative control HCC cell lines (P < 0.05).

CONCLUSIONS

CCNB1, CDK1, and RRM2 were enriched in the p53 signaling pathway and could be potential biomarkers and therapeutic targets for HBV-related HCC.

摘要

背景

肝细胞癌（HCC）是一种常见癌症，主要病因是持续性乙型肝炎病毒（HBV）感染。我们旨在确定一些与HBV相关的HCC的核心基因和通路。

方法

可从基因表达综合数据库（GEO）获取GSE62232、GSE121248和GSE94660的基因表达谱。GSE62232和GSE121248的谱图为分析数据集，GSE94660为验证数据集。应用GEO2R在线工具和韦恩图软件分析HBV相关HCC组织与正常组织之间的常见差异表达基因。然后，使用基因本体论（GO）、京都基因与基因组百科全书（KEGG）以及蛋白质-蛋白质相互作用（PPI）网络进行功能富集分析。通过Kaplan-Meier绘图仪和基因表达谱交互分析（GEPIA）检测总体生存率和表达水平。接下来，进行基因集富集分析（GSEA）以验证KEGG通路分析。此外，分别对12例HBV相关HCC患者的肿瘤组织和相邻非肿瘤肝组织、HBV相关肝癌细胞系和正常肝细胞系以及p53基因敲低或缺失的HepG2进行定量逆转录聚合酶链反应（qRT-PCR），以验证这三个核心基因的水平。

结果

选择了15个与预后明显较差相关的高表达基因，并确定p53信号通路中的CCNB1、CDK1和RRM2为核心基因。GSEA结果显示，在验证数据集中，高表达这三个核心基因的样本均富集于p53信号通路（P < 0.0001）。这三个核心基因在肿瘤组织样本中的表达高于相关相邻非肿瘤肝组织（P < 0.0001）。此外，我们还发现与正常肝细胞相比，上述基因在肝癌细胞系中高表达。另外，我们发现这三个核心基因在p53基因敲低或敲除的HCC细胞系中的表达低于阴性对照HCC细胞系（P < 0.05）。