论文部分内容阅读
目的 筛选腹主动脉瘤发生过程中与正常组织差异表达的细胞周期和细胞凋亡相关基因并作初步功能分析。 方法 应用Trizol一步法抽提 2块腹主动脉瘤壁及 2块正常主动脉组织总RNA后 ,OligotexmRNASpin Column操作法分离纯化组织mRNA ;经逆转录合成荧光分子 (Cy3/Cy5)标记cDNA探针 ,与微矩阵排列的含有 4 0 96种cDNA基因的表达谱芯片杂交 ,利用GenePixPro 3 0软件分析动脉瘤与正常组织中差异表达的基因 ,对所获得的基因进行分子生物信息学分析。 结果 腹主动脉瘤与正常组织间差异表达的细胞周期和细胞凋亡基因有 1 8条 ,占芯片基因总数的 0 44%。其中与细胞周期相关的基因 1 1条 ,与细胞凋亡相关的基因 7条。在动脉瘤组织中表达上调的基因 9条 ,平均Ratio值为 3 860 ,表达下调的 9条 ,平均Ratio值为 0 2 94。生物信息学分析显示 ,该 1 8条差异表达细胞周期和细胞凋亡基因与腹主动脉瘤中平滑肌细胞的生长和凋亡有关。 结论 腹主动脉瘤的发生、发展过程中存在多基因表达调控的改变 ,细胞周期和细胞凋亡基因与腹主动脉瘤中平滑肌细胞等的生长和凋亡有相关性 ,对于该相关基因群的进一步研究有助于阐释腹主动脉瘤的发病机制
Objective To screen the differentially expressed genes related to cell cycle and apoptosis during normal abdominal aortic aneurysm (AAAA) and analyze their primary functions. Methods Total RNA was extracted from two abdominal aortic aneurysm walls and two normal aortic tissues by Trizol in one step. Oligotex mRNA Spin Column was used to separate and purify tissue mRNA. CDNA probe was labeled with fluorescent molecule (Cy3 / Cy5) The microarray hybridization was carried out with microarray arrayed microarray containing 4096 cDNA genes. GenePixPro 30 software was used to analyze differentially expressed genes in aneurysms and normal tissues, and molecular bioinformatics analysis was performed on the obtained genes. Results There were 18 cell cycle and apoptosis genes differentially expressed between abdominal aortic aneurysm and normal tissue, accounting for 44.4% of the total number of genes in the chip. Among them, there are 11 genes related to cell cycle and 7 genes related to apoptosis. There were 9 genes that were up-regulated in aneurysm tissues, with an average Ratio value of 3860 and 9 down-regulated genes with an average Ratio value of 0 2 94. Bioinformatics analysis showed that the 18 differentially expressed cell cycle and apoptosis genes were related to the growth and apoptosis of smooth muscle cells in abdominal aortic aneurysms. Conclusion The occurrence and development of abdominal aortic aneurysm may be related to the regulation of multi-gene expression, the cell cycle and apoptosis genes and the growth and apoptosis of smooth muscle cells in abdominal aortic aneurysms. Further study will help explain the pathogenesis of abdominal aortic aneurysm