多发性硬化免疫相关基因表达谱的基因芯片研究

Immune-related gene expression profile in multiple sclerosis with gene chips

目的 用免疫相关基因芯片寻找多发性硬化表达差异的免疫相关基因。方法 将 484条免疫相关基因的double(双点 )cDNA产物按微矩阵排列点样于玻片上。例 1患者及其对照为第 1组 ,例 2患者及其对照为第 2组 ,例 3患者及其对照为第 3组。取各组外周血 ,分离淋巴细胞 ,提取总RNA ,用RT PCR逆转录成cDNA。标记cDNA探针 ,分别用cy3 dUTP、cy5 dUTP标记正常人和多发性硬化患者cDNA。将基因芯片和杂交探针变性后杂交、洗干。用ScanArray 40 0 0扫描芯片 ,GenePixPro3 0软件分析cy3、cy5两种荧光信号的强度和比值。结果 1组中筛选出差异表达的基因 2 2项 ,2组中出现差异表达的基因 2 7项 ,3组中出现差异表达的基因 72项。具有同一GeneID号的一对Double基因 ,其中 1组 6对 ,2组 9对 ,3组 30对。结论 试验结果显示正常人与MS患者免疫相关基因的表达有显著差异 ;不同发病阶段免疫相关基因表达有差异 ;以细胞免疫相关基因变化为主 。

Objective To construct the immune genes expression profile database of multiple sclerosis (MS) and to screen the high immune related candidate genes by using microarray analysis Methods Polymerase chain reaction (PCR) products on behalf of 484 immune related genes from mixed tissue library were assembled on the modified Oako glass slide The general RNA from MS patients and normal persons lymphocytes was transcripted to cDNA by RT PCR and labelled with cy 3 and cy 5 The two probes were mixed and hybridized with the above mentioned gene chips Scan array 4000 was used for scanning the hybridizing signals and GenePix Pro3 0 for date analysis Results Of the total 484 double genes monitored,22 genes of group one showed changes in expression level of the ratio outside 0 5 and 2.0;27 genes differ in group 2;72 genes change in group 3 With the same consistent gene ID of the double genes, the group one was 6,the group tow was 9 and the group three was 30 Conclusions These results show that the different expression of immune related genes might occur between MS patients and normal persons It might also afford a view of the changes that these genes should be probably related to the occurrence, development or progress of MS The cell immune related genes might be the important and most immune related genes involved in MS pathogenesis The results suggest the deeper insights into the mechanism,relapsing and treatment of MS