利用微阵列芯片技术筛选主动脉夹层致病相关基因

目的 应用微阵列芯片技术观察主动脉夹层(Stanford B型)基因表达谱的变化,筛选差异表达基因,寻找致病相关基因.方法 选取急性期的主动脉夹层(AD)标本6例,以6例正常胸主动脉作对照.抽提标本总RNA,纯化mRNA后逆转录,制备杂交探针,采用Phalanx人类全基因组芯片进行检测,应用DAVID、Pathway studio 5.0、SAM 3.01等软件对微阵列芯片的结果进行统计学、聚类和差异表达分析.选取两组间差异表达的7个基因,采用Real time PCR验证微阵列芯片结果.结果 共得到6375个基因的有效数据,其中670个差异表达基因,AD组218个基因发生上调,452个基因发生下调;其中14个基因参与细胞间黏附;12个基因参与细胞外基质的生成;4个基因参与细胞与细胞外基质间黏附;14个基因参与细胞外骨架的组成;10个基因参与免疫与炎症反应;10个基因参与胶原合成;12个基因参与细胞凋亡.Real time PCR验证结果和微阵列芯片所得数据一致.结论 AD的发生伴随着多个基因的差异表达,微阵列芯片筛选差异表达基因,有助于找到与AD致病相关的基因;上述基因的差异表达可能是AD的重要发病机制.

The screening and identification of pathogenic related genes of aortic dissection

Objective To screen the differentially expressed genes between the aorta from acute type B aortic dissections with those from normal controls and search the genes related to the pathogenesis of this disease.Methods From Sep.2005 to Feb.2008,six descending aorta specimens from patients with a-cute Stanford B dissection were taken during surgery;six normal descending aorta specimens were gotten from multi-organ donors.The gene expression profiles between two groups were compared with 6 pieces of Phalanx Whote Genome Micmarray.The data were analyzed with several professional data processing software and ouline websites.Quantitative real time pelymerase chsin reaction(Real time PCR)Was used to check the reliability of this data.Results Totally 6375 genes were valid in this experiment.Of those,670 genes were difierentially expressed in dissected and control aorta.21 8 ones were up-regulated.and 452 ones were down.regulated.Fourteen ones were related to cell.eell adhesion:12 genes to the generation of extra-cellular matrix components;4 genes to cell-matrix adhesion;14 genes to cytoskeleton formation;10 genes to immune and inflammatory response;10 genes to collagen production;12 genes to cell apoptosis.The genes including PKD1,PKD2,SPP1,COL1A1,COL3A1.COL6A2 and ACTC were verified by Real time PCR test.Conclusion The result demonstrated the complexity of the dissecting process on a molecular level.Some of these difierentially expressed genes probably resulted in the development of aortic dissection finally.