用cDNA基因芯片技术分析原发性干燥综合征免疫细胞及信号相关基因的表达

Cell and Signal Relevant Gene Expression Profile of Peripheral Blood Mononuclear Cell in Patients with Primary Sjgren's Syndrome

目的为探讨原发性干燥综合征的发病机制,应用cDNA基因芯片技术分析原发性干燥综合征患者外周血单个核细胞的免疫细胞及信号相关差异表达基因。方法Ficoll密度梯度离心法分离3例初发原发性干燥综合征患者及同年龄正常对照者外周血单个核细胞,Trizol一步法提取mRNA,反转录至cDNA,分别用Cy5及Cy3荧光标记cDNA,与寡核苷酸片段进行杂交,采用GenePix Pro 3.0图像分析软件把图像信号转化为数字信号,计算Cy5和Cy3两种荧光信号比率(ratio)。将有3个以上重复ratio的基因输入SAM系统,分析有统计学意义的免疫细胞及信号相关差异表达基因及其疾病通路(pathway)。结果原发性干燥综合征患者与正常对照者外周血单个核细胞以自然杀伤细胞介导的细胞毒通路(CD244、FASLG、CD247、GZMB、KLRD1、ICAM1)、B细胞受体信号通路(CD79A、CD19、BLNK、CD72)、细胞凋亡通路(FASLG、PRKAR2B、FADD)中的免疫细胞相关通路及基因差异最为显著(P<0.05),以上通路中有5个表达基因上调,7个表达基因下调。结论原发性干燥综合征患者的免疫细胞及信号相关通路有较多的差异表达基因,这些基因的筛选和进一步验证将为该病的发病机制及诊治提供新的思路。

Objective To explore the cell and signal relevant gene expression signatures in peripheral blood mononuclear cell （PBMC） among primary Sjogren＇s syndrome patients. Methods Three patients with primary Sjogren＇s syndrome and 3 controls were included. The cRNA probes prepared for totle RNA were hybridized with 3 identical gene chips. The difference of cell and signal relevant gene expression were compared. Results Statistical differences were calculated between patients and controls in the following three pathways： natural killer cell mediated cytotoxicity pathway, including CD244, FASLG, CD247, GZMB, KLRD1, and ICAM1, B cell receptor signaling pathway, including CD79A, CD19, BLNK, and CD72, apoptosis pathway, including FASLG, PRKAR2B, and FADD. Among these, 5 genes were upregnlated and 7 genes were downregulated. Conclusion We have identified distinct cell and signal relevant gene expression profiles of PBMC from patients with primary Sjogren＇s syndrome and health controls, along with the research advance of gene, a new moleeularbiological strategy dealing with primary Sjogren＇s syndrome will probably emerge.