基于RNA-seq筛选NSUN2调控的A549细胞差异表达基因

Screening of differentially expressed genes in A549 cells regulated by NSUN2 based on RNA-seq

检测了NSUN2在非小细胞肺癌细胞中导致的基因表达变化,可为深入研究NSUN2介导肺癌发生发展的分子机制提供更多生物学依据。采用慢病毒干扰技术,构建了稳定干扰NSUN2基因的A549细胞株,鉴定了干扰效率。以无关序列组细胞为对照,采用转录组测序技术对干扰细胞株基因表达谱进行分析,进一步筛选了差异表达基因(DEGs)。通过GO分析、KEGG pathway和GSEA富集分析,探讨了NSUN2调控的DEGs参与的生物学过程及相关通路。结果表明,与对照组细胞相比,在NSUN2低表达的A549细胞中共有1236种DEGs,其中上调基因有957个,下调基因有279个;GO富集分析发现,DEGs主要参与细胞信号传递、外部刺激反应、细胞粘着、离子通道等多个生物学过程;GSEA富集和KEGG富集分析共同显示,NSUN2干扰表达后,差异基因主要富集在整合素信号途径;进一步筛选出与整合素信号途径相关的3个关键基因(ITGβ1、ITGα5和ITGα9),RT-qPCR验证其表达水平与测序结果一致。

To detect the changes of gene expression induced by NSUN2 in non-small cell lung cancer cells,and to provide more biological basis for further study of the molecular mechanism of NSUN2-mediated carcinogenesis and development of lung cancer,Lentivirus interference technique was used to construct A549 cell line with stable interference of NSUN2 gene,and the interference efficiency was evaluated.The gene expression profiles of the two groups were detected by transcriptome sequencing technology,and the differential expressed genes(DEGs)were further screened.The biological processes and related pathways of DEGs regulated by NSUN2 were discussed by GO analysis,KEGGpathway and GSEA enrichment analysis.The results showed that 1236 DEGs are screened from NSUN2-interfering lung cancer cells,of which 957 are up-regulated and 279 are down-regulated compared with control cells;GO enrichment analysis showed that DEGS is related to biological processes such as cell signal transduction,external stimulus response,cell adhesion and ion channel;GSEA enrichment and KEGG enrichment analysis showed that after NSUN2 interference expression,the differential genes were mainly enriched in the integrin signal pathway;Three key genes related to integrin signaling pathway(ITGβ1、ITGα5 and ITGα9)have further been screened out,and their expression levels are confirmed by RT-qPCR to be consistent with the sequencing results.