摘要
目的:筛查胸腺组织重症肌无力(MG)相关基因,探讨MG发生机制.方法:分别抽提6例MG患者胸腺组织及正常胸腺组织总RNA,逆转录合成相应以Cy5和Cy3标记的cDNA一链做探针,在含有14 000点人类基因组芯片BiostarH-140 s上进行杂交,ScanArray 4000扫描仪扫描芯片荧光信号图像,用GenePix Pro3.0软件对扫描图像进行数字化处理和分析.利用实时荧光定量PCR技术,对筛选出的部分差异表达基因变化水平,在39例MG胸腺组,10例正常对照胸腺组中进行分析验证.结果:筛选出差异表达的基因254个,其中MG胸腺组中表达上调的基因82个,表达下调的基因172个.这些异常表达的基因按照功能,可以分为原癌基因和抑癌基因(3/3,即上调基因3个/下调基因3个,下同)、免疫相关蛋白(1/9)、细胞受体(1/2)、离子通道和运输蛋白(2/3)、细胞信号和传递蛋白(8/13)以及未知功能基因.验证实验显示,MG胸腺组天冬氨酰氨基葡糖苷酶(AGA)和分拣微管连接蛋白17(SNX17)相对表达量分别为[(0.671±0.078),(0.698±0.085),n=39],与正常胸腺组的[(0.611±0.043),(0.617±0.068),n=10]相比较,差异有统计学意义(P<0.05).结果与基因芯片一致.结论:MG患者胸腺组织与正常胸腺组织之间存在明显的基因表达差异,为MG患者提供相当数量的与免疫、细胞受体和细胞信号传递等相关的差异表达基因,为MG早期诊断和治疗提供了线索.
AIM： To screen the related genes for myasthenia gravis（MG） and to explore the pathogenic mechanism of MG. METHODS： Total RNA was extracted and purified from thymic tissues of 6 MG cases and 6 normal cases. The cDNAs retro-tran- scribed from RNA were labeled with Cy5 and Cy3 fluorescence as probes. The probes were hybridized with a piece of Biostar H-140 s single dot human whole gene chip. The chip was scanned with ScanArray 4000 and the acquired imaging was analyzed with Genepix Pro 3.0 software. Parts of these genes were analyzed and identified by real-time fluorescence quantitative polymerase chain reaction（FQ-PCR）. RESULTS ： Of the 254 genes with altered expressions, 82 genes were up-regulated and 172 genes were down-regulated. These genes including proto-oncogene and anti-oncogene （ 3/3, 3 up-regulated and 3 down-regulated, same as follow）, immune-associated proteinum（ 1/9）, cell receptor（ 1/ 2）, ion channel and transport protein （2/3）, and kyto-signal and transfer protein（8/13 ）. The relative expression levels of aspartyl- glyeosaminidase（AGA） and sorting nexins 17 （ SNX17 ） in thymic tissue with MG were 0. 671 ± 0. 078 and 0. 698 ＋ 0. 085. Com- pared with those in normal thymic tissues, the expression levels of AGA and SNX17 were significantly higher （ P 〈 0. 05 ）. The results consistent with those of gene chip. CONCLUSION： Marked difference exists in gene expression in thymic tissues of MG patients and normal thymic tissues, showing the complex alterations of gene expression underlying the development of MG. The identification of specific genes is helpful for early diagnosis and treatment of MG.
出处
《第四军医大学学报》
CAS
北大核心
2009年第5期454-457,共4页
Journal of the Fourth Military Medical University
基金
河南省2002年度杰出青年基金(0212000100)
河南省科研院所专项基金(0441130303)
关键词
重症肌无力
胸腺组织
基因表达谱
基因芯片
实时荧光定量PCR
myasthenia gravis
thymic tissue
gene expression profile
gene chip
Real-time fluorescence quanti- tative polymerase chain reaction
