• Zi-Wen Liang, Xiang-Dong Luo, and Zong-Cheng Yang.
• Burn Institute of Southwest Hospital, Third Military Medical University, Chongqing, 400038, China. liangzw@mail.tmmu.com.cn
• Chin. J. Traumatol. 2003 Apr 1; 6 (2): 107-13.

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.

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