Cloning of differentially expressed genes following lipopolysaccharide stimulation in human umbilical vein endothelial cells.
OBJECTIVE
To clone the differentially expressed genes in human umbilical vein endothelial cells (HUVEC) stimulated by lipopolysaccharide (LPS).


METHODS
Two-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.


RESULTS
These 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.


CONCLUSIONS
SSH is a powerful technique of high sensitivity for the detection of differential gene expression in HUVEC stimulated by LPS.
Liang
Zi-Wen
ZW
Burn Institute of Southwest Hospital, Third Military Medical University, Chongqing, 400038, China. liangzw@mail.tmmu.com.cn
Luo
Xiang-Dong
XD
Yang
Zong-Cheng
ZC
D002478:Cells, Cultured; D003001:Cloning, Molecular / Q000379:methods*; D018076:DNA, Complementary / Q000235:genetics; D004730:Endothelium, Vascular / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology; D009693:Nucleic Acid Hybridization / Q000379:methods; D016133:Polymerase Chain Reaction; D014471:Umbilical Veins / Q000378:metabolism*
D018076:DNA, Complementary; D008070:Lipopolysaccharides