肝癌细胞系PLC/PRF/5中染色质重塑蛋白ATRX表达的调控作用

Transcriptional regulation of ATRX in liver cancer cell line PLC/PRF/5

目的探讨肝癌细胞中地中海贫血伴精神发育迟滞综合征基因(ATRX)对基因表达的影响及其可能机制。方法利用CRISPR/Cas9技术敲除PLC/PRF/5细胞中的ATRX。全转录组测序技术(RNA-seq)检测ATRX敲除前后的基因表达水平,并用DESeq2软件得出差异表达基因。最后利用转座酶可及性核染色质区域分析技术(ATAC-seq)对差异基因的染色质可及性进行分析。结果分析得到2754个差异表达基因,其中MUC16和CXCL8均在ATRX缺失后表达显著上调(P<0.05),且两基因附近都存在升高的染色质可及性的区域。结论在肝癌细胞中,ATRX调控大量基因的表达,且可能直接参与了MUC16与CXCL8的抑制,为进一步研究ATRX在肝癌发生中的作用提供了新的线索。

Objective To explore the role of ATRX in gene expression with a liver cancer cell line. MethodsCRISPR/Cas9 system was employed to construct a ATRX knockout cell line. Gene expression data was generated by RNA-seq, and a differential gene expression analysis was performed using DESeq2. At last, ATAC-seq was used to assess the chromatin accessibilities of differentially expressed gene. Results Totally 2 754 genes with differential expression level were identified. Both MUC16 and CXCL8 had increased chromatin accessibilities and were significantly up-regulated(P<0.05) in the absence of ATRX. Conclusions ATRX regulates a large number of genes in liver cancer cells,so is potentially contribute to the direct inhibition of MUC16 and CXCL8.