NOX1 gene luciferase reporter gene system to establish.doc

 PAGE \* MERGEFORMAT 11 NOX1 gene luciferase reporter gene system to establish Of: Liu Zhen Liu Liu Xinghai Huang Ningwu Qi Wang Boyao [Abstract] Objective: The inflammatory cytokines TNF @, and IFN γ stimulation, the human NOX1 gene expression and regulation of a preliminary analysis. Methods: NOX1 gene 5 ‘upstream sequence connected to the non-promoter PGL3 BASIC plasmid was constructed PGL3 BASIC/NOX1 reporter plasmid. PGL3 BASIC/NOX1 A549 cells transfected with TNF @, IFN γ stimulation of 12h, dual-luciferase reporter gene system for detecting gene expression. Results: NOX1 cloned fragments with strong promoter activity , and IFN γ in the TNF @ co-stimulation, reporter gene transfection of A549 cells with the luciferase activity increased significantly compared to the control (about 4.3 times.) analysis showed that NOX1 gene 5 ‘upstream sequence fragment containing NF κB binding sites, suggesting that cytokines increased expression of luciferase may be related to activation of NF κB sites. Conclusion: NOX1 gene expression was regulated by inflammatory cytokines, suggesting that the gene may be involved in immune defense (especially the epithelial cells immune defense), it is worth further studying. [Keywords:] NOX1; reporter gene; TNF @; IFN γ The reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (NADPH oxidase, NOX) is a multi-protein subunits of the complex, expressed in phagocytes and non-phagocytic cells, can produce against pathogenic microorganisms ROS (reactive oxygen species, ROS), and participate in intracellular signal transduction, inflammation and cell proliferation processes [1]. in phagocytic cells (neutrophils, monocytes) expression was mainly NADPH oxidase gp91 phox (NOX2). NOX1 is a recent discovery of the gp91 phox homologues, in non-phagocytic cells (epithelial cells, endothelial cells) expression, as increased expression of the gene, the cells produce more ROS, and cell proliferation and other cell behavi