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Effects of Heterologous Expression of Thioredoxin Reductase on the Level of Reactive Oxygen Species in COS-7 Cells  

Kang, Hyun-Jung (Division of Life Sciences, College of Natural Sciences, Kangwon National University)
Hong, Sung-Min (Division of Life Sciences, College of Natural Sciences, Kangwon National University)
Kim, Byung-Chul (Division of Life Sciences, College of Natural Sciences, Kangwon National University)
Park, Eun-Hee (College of Pharmacy, Sookmyung Women's University)
Ahn, Kisup (Department of Health and Environment, Baekseok College of Cultural Studies)
Lim, Chang-Jin (Division of Life Sciences, College of Natural Sciences, Kangwon National University)
Abstract
Thioredoxin reductase (TrxR), a component of the redox control system involving thioredoxin (Trx), is implicated in defense against oxidative stress, control of cell growth and proliferation, and regulation of apoptosis. In the present study a stable transfectant was made by introducing the vector pcDNA3.0 harboring the fission yeast TrxR gene into COS-7 African green monkey kidney fibroblast cells. The exogenous TrxR gene led to an increase in TrxR activity of up to 3.2-fold but did not affect glutathione (GSH) content, or glutaredoxin and caspase-3 activities. Levels of reactive oxygen species (ROS), but not those of nitric oxide (NO), were reduced. Conversely, 1-chloro-2,4-dinitrobezene (CDNB), an irreversible inhibitor of mammalian TrxR, enhanced ROS levels in the COS-7 cells. After treatment with hydrogen peroxide, the level of intracellular ROS was lower in the transfectants than in the vector control cells. These results confirm that TrxR is a crucial determinant of the level of cellular ROS during oxidative stress as well as in the normal state.
Keywords
COS-7; Hydrogen Peroxide; Reactive Oxygen Species; Thioredoxin Reductase;
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