猪内皮细胞与血清作用后表达上调基因的初步鉴定

A Preliminary Identification of Up-regulated Genes on Porcine Endothe-lial Cells by Human Serum Activation

目的探讨猪内皮细胞与人血清作用后基因表达的变化。方法猪内皮细胞系PIEC被含 2 0 %人血清的培养基作用 2 .5小时作为实验组 ,对照组为含 2 0 %胎牛血清的培养基作用的PIEC。利用抑制消减杂交技术 (SSH)、DNA序列测定技术、生物信息学分析、RT PCR等进行研究。结果经过SSH和克隆 ,获得一包含约 2 0 0个克隆的消减文库。从文库中随机挑取 15个克隆进行测序 ,并在GenBank中进行同源性比较。其中 1个是猪IL 8的部分序列 ,3个序列分别与人唐氏综合征核心区基因 1(DSCRl)、人FSHD区基因I(FRGl)、牛伴侣蛋白 10cpnlo基因同源。其余序列尚无已知同源基因匹配。结论猪内皮细胞与人血清作用后有许多基因的表达上调。

Objective To investigate gene expression change on human serum activated--porcine endothelial cells. Methods Porcine vascular endothelial cell line PIECs were treated for 2.5 hours with 20% human sera. The control is 20% fetal calf serum (FCS) treating PIECs. Suppression subtraetive hybridization (SSH), DNA sequencing, bioinformatics analysis, RT-PCR were used. Results By means of SSH and cloning, we obtained a subtracted cDNA library which included about 200 clones corresponding to up-regulated genes. 15 of these clones were identified by sequencing and analyzed for homology in the GenBank. 1 of 15 is part of porcine IL-8 gene. Three of the sequences have similar homology with human Down syndrome critical region gene 1 (DSCR1), human FSHD region gene 1 (FRG1) and bovine chaperonin 10 (cpnl0). The rest were no database match.Conclusion Many genes were up-regulated on human serum activated-porcine endothelial cells.