人血管内皮细胞缺氧早期基因系列表达分析文库的建立及分析

Establishment and Gross Analysis of Serial Analysis of Gene Expression Libraries of Human Vascular Endothelial Cell before and after Hypoxia

目的:建立常氧及缺氧早期人血管内皮细胞基因系列表达分析文库,对表达谱数据进行初步分析,为创伤后多器官功能障碍的防治奠定基础。方法:体外培养人脐静脉内皮细胞株(EA.hy926),分为正常对照组和缺氧3h处理组。采用长标签基因系列表达分析方法(LongSAGE)建立常氧及缺氧3h组人血管内皮细胞基因表达文库,通过与Genebank数据库比对确认基因,对所得标签进行初步分析。结果:通过LongSAGE技术构建了常氧和缺氧脐静脉内皮细胞LongSAGE文库。缺氧SAGE文库测得30756个标签,其中识别出的基因总数为13879,比例为45.13%;重复的双标签体总数为762。常氧SAGE文库测得标签数为31213,其中识别出的基因总数为13665,比例为43.78%;重复的双标签体总数为758。结论:成功建立了人脐静脉内皮细胞常氧和缺氧早期LongSAGE文库,获得涵盖上万个转录本信息的表达谱数据,为创伤后多器官功能障碍的防治奠定基础.

Objective： To establish Long SAGE libraries of human vascular endothelial cells before and early after hypoxia, and priliminarily analyse the data so as to make basis for the management of multiple organ dysfunction after trauma. Methods： Human umbilical endothelial cell line （EA.hy926） was divided into normal control and 3 hrs of hypoxia treatment groups. After the establishment of Long SAGE libraries of normoxia and hypoxia endothelial cells, the colonies were screened by PCR. Then the SAGE tags were extracted for gross analysis. Results we successfully constructed the Long SAGE libraries of normal and hypoxic umbilical endothelial cells. Totally 30756 tags in hypoxic SAGE library, among which the unique tags were 13879 （accounting for 45.13 %）, and the repeated Ditags were 762. In the normoxic library, there were 31213 tags were attained,among which the unique tags were 13665 （accounting for 43.78 %）, and the repeated Ditags were 758. Conclusion： Gene expression profile covering tens of thousands of transcripts was bulild, which provides basis for the analysis of changes in vascular endothelial cells early after hypoxia.