用基因表达谱芯片筛选原发性下肢静脉曲张相关基因的研究

A study on differently expressed genes in primary varicose vein by cDNA microarray

目的运用基因表达谱芯片研究原发性下肢静脉曲张基因表达谱的变化。方法选取原发性下肢静脉曲张患者隐 股交界瓣膜区静脉 5条 ,取 5条正常静脉作对照。抽提总RNA、纯化mRNA、反转录制备杂交探针 ,应用含有 4 0 96种人类基因全长cDNA的表达谱芯片进行差异表达谱分析 ,随后应用反转录PCR验证部分差异表达基因。结果曲张大隐静脉瓣膜区表达谱中差异表达基因共有 16 8条 ,上调基因 96条 ,下调基因 72条 ;5例标本均差异表达的基因共 39条 ,其中上调2 8条 ,下调 11条 ;多条细胞凋亡相关基因、细胞信号和传递蛋白基因、原癌基因和抑癌基因等均有差异表达 ;反转录PCR证实caspase 9及MAP3K基因及其蛋白产物在曲张静脉中差异表达。 结论应用基因表达谱芯片筛选致病相关基因 ,可能为下肢静脉曲张的发病机制研究提供新思路。

Objective To screen the differently expressed genes in great saphenous varicose vein using cDNA microarray. Methods Varicose veins from 5 cases and normal veins near the saphenofemoral junction were collected, total RNA was isolated and mRNA was purified by oligotex. mRNA from varicose veins and normal veins were reversely transcribed to cDNA with the incorporation of fluorescent dUTP to prepare the hybridization probes, which were hybridized to cDNA microarray of 4 096 human genes. RT-PCR and Western blot were used to identify differently expressed genes. Results One hundred and sixty-eight genes were shown in differently expressed profile with 96 upregulated and 72 downregulated genes, in all 5 varicose samples there were 28 genes upregulated and 11 downregulated. The differently expressed genes were involved in the functions of apoptosis, signal conduct, protooncogene and anti-oncogene. The expression of caspase 9 and MAP3K were identified by RT-PCR and Western blot. Conclusion Disease-related gene screening by cDNA microarray in great saphenous varicose vein affords an insight into the pathogenesis of varicosis.