肺栓塞患者血浆凝血因子基因表达谱的差异  

Difference in gene expression profile of plasma coagulation factors in patients with pulmonary embolism

作　　者：程克斌[1] 王乐民[2] 胡莺[3] 郜恒骏[3] 

机构地区：[1]同济大学附属肺科医院呼吸科,上海200433 [2]同济大学附属同济医院心内科 [3]生物芯片上海国家工程研究中心

出　　处：《上海医学》2008年第6期381-384,共4页Shanghai Medical Journal

基　　金：国家自然科学基金(30570809)

摘　　要：目的应用基因芯片技术比较肺栓塞(PE)患者与健康者凝血因子基因表达谱的差异,探讨血浆中凝血因子mRNA水平与PE的关系。方法取9例PE患者(PE组)和33名健康者(正常对照组)的静脉血5 mL,抽提血液总RNA。合成9个Cy3标记的PE患者cRNA探针和1个Cy5标记的标准对照探针,与44 000 Agilent人全基因组Oligo芯片杂交,用Agilent扫描仪扫描荧光信号。应用Feature Extraction软件分析、计算每点的Cy3和Cy5信号值。以9组数据中,Cy3/Cy5比值(Ratio比值)为标准,筛选差异表达基因。并采用荧光定量-聚合酶链反应进行验证。结果9张芯片筛选出共同差异表达的凝血因子基因13条,其中mRNA表达水平上调基因11条,分别为FⅦ、FⅪ、FⅧA1、FⅤ、FⅧ、FⅩ、FⅡ、vWF、FGG、F2R、F2RL1;表达下调基因2条,分别为FⅨ和FⅫ。结论血浆凝血因子的差异表达可能与PE的发病有关。Objective To identify the association between the mRNA expression of blood coagulation factors and pulmonary embolism （PE） by comparing the gene expression profiles of patients with PE and normal controls using gene microarray. Methods The fresh venous blood samples were collected from 9 PE patients and 33 normal controls, and the total RNA was extracted and purified. We prepared 9 PE cRNA probes labeled with cyanine 3 and a standard cRNA probe labeled with cyanine 5, which were used for hybridization with Agilent Whole Human Genome Oligo Microarray. Then the chips were scanned by Agilent scanner and the acquired image was analyzed by Feature Extraction software to compute the ratio of Cy3 to CyS. The differentially expressed genes were identified if the value of the ratio of Cy3-CTP to CyS-CTP was more or less than 1. The result of chips was verified by fluorescence quantitative polymerase chain reaction （FQ-PCR）. Results Eleven genes of them were randomly selected to be amplified by FQ-PCR. The differential expression has been observed in all the selected genes, which is consistent with the microarray. Thirteen differentially expressed genes of blood coagulation factors were screened from the 9 microarrays. There were 11 genes upregulated, including FⅦ, FⅪ, FⅫAl, FV, FⅦ, FⅩ, FⅡ, vWF, FGG, F2R, and F2RL1; and these were 2 genes downregulated, including F Ⅸ and F Ⅻ. Conclusion Differential expression of blood coagulation factor genes in the blood may play a role in the initiation and development of PE.

关 键 词：肺栓塞 遗传学 基因芯片 荧光定量-聚合酶链反应 凝血因子 

分 类 号：R563.5[医药卫生—呼吸系统]