应用基因芯片筛选慢性乙型肝炎患者PD-1／PD—L通路基因差异表达

Screening the differentially expressed genes in PD-1/PD-L signal pathways in chronic hepatitis B（ CHB ） patients by gene expression microarray

目的应用基因芯片技术筛选慢乙肝病人PD-1／PD—L通路差异性表达的基因。方法应用基因芯片技术筛选4例慢乙肝病人与2例健康人外周血，获得差异性表达的基因，结合初步构建的PD-1／PD-L信号通路，挑选出_致的转录因子或调控蛋白，并应用Westernblot方法检测CD40L、Bcl-10、GATA-3、NFAT蛋白的水平的表达。结果与健康人相比，慢乙肝患者有838个异常表达的基因，其中高表达的基因有150个，低表达的基因有688个。在这些异常表达基因中，有13个基因属于所构建的PD．1／PD．L信号通路中的转录因子或调控蛋白，其中表达上调的6个分别是BCL-10、AP、SRY、MYOD1、ELK4、NFAT，表达下调的7个分别是CD40L、GADS、PDK1、IL4、GATA3、CREB、RARG。Westem blotting法证实慢乙肝组高表达BCL-10，低表达CD40L、GATA3，NFAT但与病毒载量不相关。结论CIMOL、BCL-10、GATA3、NFAT在慢性乙型肝炎患者PD-1／PD—L信号传导通路中可能有重要的调控作用，可以做深入的功能分析研究。

Objective To screen the gene expression profiles in PD-1/PD-L signal pathways in chronic hepatitis B（ CHB ）patients by gene expression microarray . Methods The differentially expressed genes in peripheral blood samples of 4 cases with CHB and 2 normal people was determined by Gene expression microarray, and then,in combination with prehminary construction of PD-1/PD-L signal pathways, selecting the consistent transcription factors or regulatory protein. The CD40L,BCL-10,GATA3 and NFAT protein expression were detected by Western blot. Results 838 differentially expressed genes were detected, 150 genes were up- regulated and 688 genes were down-regulated. 13 genes are belonging to the transcription factors or regulatory protein of PD-1/PD-L signaling pathways that we have constructed. In these 13 genes,6 genes （BCL-IO,AP, SRY, MYOD1, ELK4, NFAT） are down-regulated and 7 genes （ CD 40L, GADS, PDK1, IL-4, GATA3, CREB, RARG） are down-regulated . Using the Western blotting method,we found that BCL-IO protein expression was up - regulated in CHB patients, but CD 4 0 L, GATA 3 and NFAT proteins expression were down - regulated, which .was not related with viral load carrying capacity. Conclusions CD 40L, BCL-10, GATA3 and NFAT genes may have an important role in regulating the PD-1/PD-L signaling pathway. Further functional study is still need to be done.