CLONING OF LIPOPOLYSACCHARIDE RESPONSIVE GENES IN ENDOTHELIAL CELLS BY SUPPRESSION SUBTRACTIVE HYBRIDIZATION
- VernacularTitle:应用抑制消减杂交克隆内皮细胞内毒素刺激后相关基因
- Author:
Ziwen LIANG
;
Xiangdong LUO
;
Zongcheng YANG
- Publication Type:Journal Article
- Keywords:
endothelium, vascular;
endotoxins;
suppress subtractive hybridization
- From:
Medical Journal of Chinese People's Liberation Army
1981;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
To screen genes in endothelial cells resulted from responding to lipopolysaccharide (LPS), mRNA was extracted from both untreated human umbilical endothelial cells (HUVEC) following and HUVEC which were treated with LPS for 6 hours. cDNAs of both populations were synthesized to generate cDNA libraries by suppression subtractive hybridization (SSH). The libraries were then screened with colony dot blots. Positive clones were sequenced and BLAST analysed. The results showed differential genes included 3 novel genes and 22 known genes. The 3 novel genes were confirmed by Northern blotting analysis. These 22 known genes were involved in the regulation of proinflammatory response, cell apoptosis, cytoskeleton, signal transduction and energy metabolism. These results suggest that SSH is an effective technique to detect differential gene expression in HUVEC, which may be helpful to evaluate molecular mechanisms of endothelial injury induced by LPS and provide potential therapeutic targets for LPS related disturbances.
