Cloning of differentially expressed genes following lipopolysaccharide stimulation in human umbilical vein endothelial cells.
- Author:
Zi-Wen LIANG
1
;
Xiang-Dong LUO
;
Zong-Cheng YANG
Author Information
- Publication Type:Journal Article
- MeSH: Cells, Cultured; Cloning, Molecular; methods; DNA, Complementary; genetics; Endothelium, Vascular; metabolism; Gene Expression; Humans; Lipopolysaccharides; pharmacology; Nucleic Acid Hybridization; methods; Polymerase Chain Reaction; Umbilical Veins; metabolism
- From: Chinese Journal of Traumatology 2003;6(2):107-113
- CountryChina
- Language:English
-
Abstract:
OBJECTIVETo clone the differentially expressed genes in human umbilical vein endothelial cells (HUVEC) stimulated by lipopolysaccharide (LPS).
METHODSTwo-directional (forward and backward) suppression subtractive hybridization (SSH) was performed on HUVEC cultured in either standard media or treated for 6 hours with LPS (100 ng/ml). To restrict the number of false-positive clones, colony dot hybridization was used to further verify the differentially expressed cDNA clones. Positive clones were sequenced.
RESULTSThese analyses have identified both novel and known genes whose expression is influenced by LPS. The known genes include a group related to proinflammatory events, a group related to cellular apoptosis and proliferation, a group related to protein synthesis and cytoskeletal rearrangment, and a group related to energy metabolism and signal transduction.
CONCLUSIONSSSH is a powerful technique of high sensitivity for the detection of differential gene expression in HUVEC stimulated by LPS.
