基于公共数据库筛选淋巴特异性解旋酶过表达肺腺癌细胞的差异微小RNA及其关键基因对预后的影响
Differential microRNA of lymphoid specific helicase overexpression lung adenocarcinoma cell and effect of its key gene on prognosis based on public database screening
刘崇梅 1曾毅群 1许勇 1陈睿鹏 1高亚1
作者信息
- 1. 湖南师范大学附属岳阳医院病理科,湖南 岳阳 414000
- 折叠
摘要
目的 分析淋巴特异性解旋酶(LSH)在肺腺癌PC9细胞过表达后微小RNA(miRNA)的表达差异,并预测其潜在调控基因.方法 采用高通量测序技术检测LSH过表达的PC9细胞系中的总miRNA,并筛选核质比有差异的候选miRNA,采用实时荧光定量聚合酶链反应(PCR)对候选miRNA的核质比进行验证,应用生物信息学方法对调控基因进行京都基因与基因组百科全书(KEGG)富集分析及基因本位(GO)功能富集分析,构建蛋白质-蛋白质相互作用(PPI)网络对miRNA核靶点进行筛选,利用癌症基因组图谱(TCGA)数据库探究候选靶基因对肺腺癌患者预后的影响.结果 通过对LSH过表达PC9细胞进行测序及基因差异分析,共筛选出5个核质比差异的miRNA:hsa-miRNA-30e-5p、hsa-miRNA-378i、hsa-miRNA-378f、hsa-miRNA-423-5p、hsa-miRNA-4284,其中核内miRNA-4284增加最为显著.经KEGG与GO富集分析发现这些miRNA可能通过结合靶基因参与了RAS信号通路的调控.通过PPI网络及肺癌预后预测发现,差异miRNA可下调CXC趋化因子受体4(CXCR4)表达,增加肺腺癌患者的死亡风险.结论 LSH过表达的肺腺癌PC9细胞核和细胞质中出现多个显著差异表达的miR-NA,以核内miRNA-4248变化最为明显,生物信息学分析发现差异miRNA可能通过下调CXCR4的表达增加肺腺癌患者的死亡风险.
Abstract
Objective To analyze the expression difference of microRNA(miRNA)after overexpression of lymphoid specific helicase(LSH)in lung adenocarcinoma PC9 cells,and to predict its potential regulatory genes.Method The total miRNA in LSH-overexpressed PC9 cell lines were detected by high-throughput gene sequencing technology,and candi-date miRNA with different nucleoplasmic ratios were screened,and the nucleoplasmic ratios of candidate miRNA were verified by real-time quantitative polymerase chain reaction(PCR).The Kyoto Encyclopedia of Genes and Genomics(KEGG)enrichment analysis and Gene Ontology(GO)functional enrichment analysis were performed on regulatory genes by bioinformatics methods,and protein-protein interaction(PPI)networks were constructed to screen miRNA nu-clear targets.The effects of candidate target genes on the prognosis of lung adenocarcinoma patients were investigated by The Cancer Genome Atlas(TCGA)database.Result By sequencing and gene difference analysis of LSH-overexpressed PC9 cells,five miRNA with different nucleoplasmic ratio were screened:hsa-miRNA-30e-5p,hsa-miRNA-378i,hsa-miR-NA-378f,hsa-miRNA-423-5p,hsa-miRNA-4284,among which the increase of nuclear miRNA-4284 was the most signif-icant.KEGG and GO enrichment analysis showed that these miRNA might be involved in the regulation of RAS signal-ing pathway by binding target genes.Through PPI networks and lung cancer prognosis prediction,differential miRNA down-regulate the expression of C-X-C motif chemokine receptor 4(CXCR4)and increased the death risk of lung adeno-carcinoma patients.Conclusion LSH-overexpressed lung adenocarcinoma PC9 has multiple miRNA with significantly different expression in the nucleus and cytoplasm,and the change of miRNA-4248 in the nucleus is the most obvious.Bioinformatics analysis shows that different miRNA may increase the death risk of lung adenocarcinoma patients by down-regulating CXCR4 expression.
关键词
肺癌/淋巴特异性解旋酶/微小RNA/miRNA-4248/CXC趋化因子受体4Key words
lung cancer/lymphoid specific helicase/microRNA/miRNA-4248/C-X-C motif chemokine receptor 4引用本文复制引用
基金项目
湖南省自然科学基金面上项目(2022JJ30578)
出版年
2024
NETL
NSTL
万方数据