敲除ARID1A对人肝癌细胞系PLC/PRF/5基因表达调控的影响

Effects of ARID1A loss on transcriptionalregulation in hepatocellular carcinoma cell line PLC/PRF/5

目的探讨肝癌细胞系PLC/PRF/5中敲除ARID1A后对基因表达调控的影响。方法利用CRISPR-Cas9基因编辑技术敲除PLC/PRF/5细胞中ARID1A。然后分别对PLC/PRF/5 ARID1A野生型与敲除型细胞进行全转录组测序分析(RNA-seq),并用DESeq2鉴定ARID1A敲除后差异表达基因。最后,使用Metascape数据库对差异表达基因进行GO功能富集分析。结果成功构建PLC/PRF/5 ARID1A敲除细胞系。利用RNA-seq共筛选出978个差异表达基因,包括480个表达上调差异基因和498个表达下调差异基因。GO功能富集分析显示差异表达基因主要集中在细胞黏附、激素水平调节、激素代谢和MAP激酶活性调节等生物学过程。结论PLC/PRF/5肝癌细胞中,ARID1A可调控大量基因的表达,为进一步研究ARID1A在肝癌发生发展中的作用机制提供了新的线索。

Objective To explore the transcriptional alterations regulated by ARID1A in hepatocellular carcinoma cell line PLC/PRF/5.Methods CRISPR/Cas9 technology was utilized to generate ARID1A knockout cells.RNA-seq was then performed in both ARID1A intact and ARID1A null PLC/PRF/5 cells to generate gene expression profiles.Differentially expressed genes were identified using DESeq2.Functional enrichment analysis of differentially expressed genes was performed using metascape database.Results Successful generation of the ARID1A knockout PLC/PRF/5 cells was confirmed by Western blot and Sanger sequencing.A total of 978 differentially expressed genes were identified by RNA-seq,among which the expression of 480 genes was up-regulated and that of 498 genes was down-regulated.These differentially expressed genes were mainly enriched in several biological processes including cell adhesion,regulation of hormone levels,hormone metabolism and regulation of MAP kinase activity.Conclusions The results indicate that ARID1A loss induces alterations in transcription in PLC/PRF/5,which may provide novo clues for elucidating the underlying mechanism of ARID1A in tumorigenesis.