乙型肝炎病毒相关肝细胞癌中关键microRNA的鉴定及其功能研究

摘要/Abstract

摘要： 目的：乙型肝炎病毒（hepatitis B virus, HBV）相关肝细胞癌（hepatocellular carcinoma, HCC）关键microRNA的鉴定及其对HCC细胞增殖和迁移能力的影响。方法：①利用GEO2R工具分析GEO数据库中GSE69580和GSE67882数据集,筛选HBV相关HCC组织与正常肝脏组织之间的差异表达miRNAs（DEMs）;②利用miRTarBase数据库预测miRNAs的靶基因,并通过DAVID数据库对DEMs的靶基因进行基因本体论（GO）功能注释分析与京都基因和基因组百科全书（KEGG）通路富集分析;③利用蛋白-蛋白互作（PPI）网络分析确定DEMs的关键miRNAs,并利用miRNACancerMAP数据库分析miRNAs可能参与的信号通路;④使用Kaplan-Meier Plotter数据库分析关键miRNAs与HCC临床预后的关系;⑤使用MTT法和划痕实验验证DEMs对HCC细胞增殖和迁移能力的影响。结果：通过生物信息学分析共鉴定出12个上调DEMs和1个下调DEMs,并通过GO和KEGG功能富集分析表明,miR-93和miR-125b参与MAPK、PI3K-AKT和P53等肿瘤相关信号通路。通过Kaplan-Meier plotter生存分析发现,miR-93表达水平与HBV相关HCC患者的预后负相关（P=0.036）,而miR-125b表达水平与HBV相关HCC患者的预后呈正相关关系（P=0.032）。在HBsAg阳性的肝癌细胞系MHCC-97H中,过表达miR-93或抑制miR-125b均可增强MHCC-97H细胞的增殖和迁移能力（均P<0.05）,而过表达miR-125b或抑制miR-93可降低MHCC-97H细胞增殖和迁移能力（均P<0.05）。结论：miR-93和miR-125b是HBV相关HCC的关键miRNAs,且上调miR-125b或下调miR-93可抑制肝癌细胞系MHCC-97H的增殖和迁移能力。

关键词: 肝细胞癌, 微小RNA, 增殖, 预后

Abstract: Objective: To investigate the hub microRNA of hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) and its effect on the proliferation and migration ability of HCC cells. Methods: ①The GEO2R tool was used to analyze the GSE69580 and GSE67882 datasets in the GEO database to screen for differentially expressed miRNAs (DEMs) between HBV-related HCC tissues and normal liver tissues. ②The miRTarBase database was used to predict the target genes of miRNAs and the DAVID database was used to target DEMs Gene annotation analysis of gene ontology (GO) and enrichment analysis of Kyoto Gene and Genome Encyclopedia (KEGG) pathway. ③Protein-protein interaction (PPI) network analysis was used to identify hub miRNAs of DEMs and the miRNACancerMAP database was used to analyze the signal pathways that miRNAs might be involved in. ④Kaplan-Meier Plotter database was used to analyze the relationship between hub miRNAs and HCC clinical prognosis. ⑤MTT method and scratch experiments were used to verify the effect of DEMs on the proliferation and migration of HCC cells. Results: A total of 12 up-regulated DEMs and one down-regulated DEM were identified by bioinformatics analysis, and GO and KEGG functional enrichment analysis indicated that miR-93 and miR-125b were involved in tumor-associated signaling pathways such as MAPK, PI3K-AKT, and P53. Kaplan-Meier plotter survival analysis revealed that miR-93 expression level was negatively correlated with the prognosis of patients with HBV-related HCC (P=0.036), while miR-125b expression level was positively correlated with the prognosis of patients with HBV-related HCC (P=0.032). In HBsAg-positive hepatocellular carcinoma cell line MHCC-97H, overexpression of miR-93 or inhibition of miR-125b increased the proliferation and migration capacity of MHCC-97H cells (P<0.05), while overexpression of miR-125b or inhibition of miR-93 decreased the proliferation and migration capacity of MHCC-97H cells (P<0.05). Conclusion: miR-93 and miR-125b are the hub miRNAs for HBV-associated HCC, and up-regulation of miR-125b or down-regulation of miR-93 inhibited the proliferation and migration capacity of the hepatocellular carcinoma cell line MHCC-97H.

Key words: Hepatocellular carcinoma, MicroRNA, Proliferation, Prognosis

裴志燕, 王健力, 张岭漪. 乙型肝炎病毒相关肝细胞癌中关键microRNA的鉴定及其功能研究[J]. 肝癌电子杂志, 2020, 7(4): 29-37.

Pei Zhiyan, Wang Jianli, Zhang Lingyi. Identification and functional analysis of key microRNAs in HBV-associated hepatocellular carcinoma[J]. Electronic Journal of Liver Tumor, 2020, 7(4): 29-37.

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