基因表达谱芯片筛选破裂性腹主动脉瘤组织中的差异表达基因

Study on Differently Expressed Genes in Ruptured Abdominal Aorta Aneurysm by cDNA Microarray

目的采用表达谱基因芯片技术筛选破裂性腹主动脉瘤组织中的差异表达基因。方法选取破裂性腹主动脉瘤组织标本6例,以年龄、性别构成、动脉瘤直径等相仿的6例未破裂腹主动脉瘤作对照。抽提总RNA,纯化mRNA后逆转录制备杂交探针,采用含有4 096种人类基因全长cDNA的表达谱芯片进行差异表达谱分析,通过Northern blot或Western blot反向验证部分差异表达基因。结果在破裂性腹主动脉瘤组织中发现差异表达基因共有124条,上调的基因89条,下调的基因35条;共存性基因有42条,其中上调基因29条,下调基因13条。差异表达的基因有与基质代谢、炎性因子及粘附分子、细胞凋亡、原癌基因或抑癌基因、细胞信号和传递蛋白、细胞骨架和运动蛋白相关基因表达密切相关的多种基因参与。通过Northern blot等方法反向验证基质金属蛋白酶9和金属蛋白酶组织抑制剂2基因,证实在腹主动脉瘤组织中存在基质金属蛋白酶9和金属蛋白酶组织抑制剂2的mRNA及蛋白产物的差异表达。结论应用表达谱基因芯片筛选破裂性腹主动脉瘤中的差异表达基因,可能为其发病机制及防治的研究提供新思路。

Aim To screen the differently expressed genes in ruptured abdominal aorta aneurysm （RAAA） using cDNA micrearray. Methods 6 RAAA or abdominal aorta aneurysm （ AAA） specimen were collected respectively, and total RNA was isolated from the tissues and purified to mRNA by ohgotex. Both mRNA from AAA and normal aorta were reversely transcribed to cDNA with the incorporation of fluorescent dUTP to prepare the hybridization probes, then the mix probes were hybridized to cDNA micrearray of 4 096 human genes. After washing, the cDNA micrearray were scanned for the fluorescent signals and showed differences between 2 tissues. Noahem and Western blot were used to identify the differently expression of some genes in RAAA. Results Among 4 096 target genes, 124 genes were showed in differential, expressed profile with 89 upregulated and 35 downregulated genes, and all 6 AAA samples were 29 genes upregulated and 13 downregulated. The differential expressed genes involved in the functions of matrix metalloproteinases （MMP）, inflammation, apoptosis, signal conduct, proto-oncogene and anti-oncogene, which meant that they had potential value in the pathogenesis of AAA. Then, Noahem and Western blot were used to identify the different expression of MMP-9 and tissue inhibitors of metalloproteinase-2 （TIMP-2） in RAAA. Conehtsion The investigation based on cDNA microarray in researching related genes in RAAA afforded a new idea in studying pathogenesis, prevention and therapy of RAAA.