Objective:The aim of this study was to investigate whether astrocyte elevated gene 1 (AEG1) regulates COX-2 expression in human hepatoma HepG2 cells and related pathways involved in this process. Methods:Human hepatom...Objective:The aim of this study was to investigate whether astrocyte elevated gene 1 (AEG1) regulates COX-2 expression in human hepatoma HepG2 cells and related pathways involved in this process. Methods:Human hepatoma HepG2 cells were transfected with pcDNA3.1(-)-AEG1 plasmid or psilencer2.0-AEG1-shRNA1 plasmid to up/down-regulate AEG1 expression, pcDNA3.1(-) and psilencer 2.0 empty vector plasmids were transfected respectively as control. Real-time RT-PCR was carried out to measure the expression levels of AEG1 and COX-2 mRNA. The expression levels of AEG1 and COX-2 protein were detected by Western blot. NF-κB signaling was blocked by PDTC, and AP-1 signaling was blocked by curcumin. Results:AEG1 mRNA and protein levels were increased after pcDNA3.1(-)-AEG1 transfection, and decreased after psilencer2.0-AEG1-shRNAs transfection. COX-2 mRNA and protein levels were increased in AEG1-overexpressing cells and decreased in AEG1-knockdown cells. Phosphorylations of p65 and c-jun were up-regulated in AEG1-overexpressing cells. Both PDTC and curcumin reduced COX-2 expression in HepG2 cells with AEG1 overexpression. Conclusion:AEG1-overexpressing and-knockdown HepG2 cells are established successfully. AEG1 could induce COX-2 expression though activating NF-κB and AP-1 in human hepatoma HepG2 cells.展开更多
基金Supported by grants from the National Science Foundation of China (No.81070333)the Natural Science Foundation of Hubei Province of China (No.2012FFB02318)
文摘Objective:The aim of this study was to investigate whether astrocyte elevated gene 1 (AEG1) regulates COX-2 expression in human hepatoma HepG2 cells and related pathways involved in this process. Methods:Human hepatoma HepG2 cells were transfected with pcDNA3.1(-)-AEG1 plasmid or psilencer2.0-AEG1-shRNA1 plasmid to up/down-regulate AEG1 expression, pcDNA3.1(-) and psilencer 2.0 empty vector plasmids were transfected respectively as control. Real-time RT-PCR was carried out to measure the expression levels of AEG1 and COX-2 mRNA. The expression levels of AEG1 and COX-2 protein were detected by Western blot. NF-κB signaling was blocked by PDTC, and AP-1 signaling was blocked by curcumin. Results:AEG1 mRNA and protein levels were increased after pcDNA3.1(-)-AEG1 transfection, and decreased after psilencer2.0-AEG1-shRNAs transfection. COX-2 mRNA and protein levels were increased in AEG1-overexpressing cells and decreased in AEG1-knockdown cells. Phosphorylations of p65 and c-jun were up-regulated in AEG1-overexpressing cells. Both PDTC and curcumin reduced COX-2 expression in HepG2 cells with AEG1 overexpression. Conclusion:AEG1-overexpressing and-knockdown HepG2 cells are established successfully. AEG1 could induce COX-2 expression though activating NF-κB and AP-1 in human hepatoma HepG2 cells.
