摘要
目的利用基因芯片技术研究人主动脉瘤组织与正常主动脉组织基因表达谱的差异。方法选取主动脉瘤病例标本5例,正常主动脉标本4例。提取总RNA,反转录成c DNA、体外转录合成aRNA后与芯片进行杂交,对结果进行分析。同时利用RT-qPCR对从表达谱筛选出的其中6个差异基因进行基因转录水平的定量验证。结果人主动脉瘤组与正常主动脉组相比,表达差异倍数大于2的基因共有270个,其中有211个基因上调表达,59个基因下调表达。差异表达的基因主要参与信号传导、免疫反应和炎性反应等生物学过程。RT-qPCR检测结果与基因芯片结果的符合率为100%。结论主动脉瘤与正常主动脉的表达谱有较大差异,利用基因芯片技术结合RT-qPCR验证可以筛选出主动脉瘤差异表达的基因,为研究主动脉瘤发病机制提供分子基础。
Objective To investigate the difference of gene expression profile in aortic aneurysm and normal aorta.Methods Total RNA of 5 cases of aortic aneurysm and 4 normal aorta were isolated and c DNA was synthesized,then aRNA labeled by biotin was synthesized by in vitro transcription. The mixed probes were hybridized with Affymetrix Gene Chip Human Genome U133 Plus 2. 0 Array. The fluorescent signal was scanned by Affymetrix Gene Chip Scanner3000. The acquired image was analyzed by GCOSv L. 4 software. 6 different expression genes from profiles were validated by RT-qPCR. Results Totally 270 genes had more than two fold change in expression in aortic aneurysm,which included 211 upregulated and 59 downregulated genes. Different expressed genes were mainly involved in signal transduction,immune system process,and inflammatory response and so on. Six genes associated with formation of aortic aneurysms were selected for quantitative analysis,and the results of RT-qPCR were completely consistent with the results of gene microarray. Conclusions Gene differentially expressed in aortic aneurysms identified by microarray may provide a molecular basis for the research of pathogenesis of aortic aneurysm.
出处
《基础医学与临床》
CSCD
2016年第11期1478-1482,共5页
Basic and Clinical Medicine
基金
国家自然科学基金(81270385)
