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http://dx.doi.org/10.5352/JLS.2013.23.2.167

Construction and In vitro Study of a Prx 6/Luc Vector System for Screening Antioxidant Compounds in the Transgenic Mice  

Lee, Young Ju (Department of Biomaterials Science, College of Natural Resources & Life Science, Pusan National University)
Nam, So Hee (Department of Biomaterials Science, College of Natural Resources & Life Science, Pusan National University)
Kim, Ji Eun (Department of Biomaterials Science, College of Natural Resources & Life Science, Pusan National University)
Hwang, In Sik (Department of Biomaterials Science, College of Natural Resources & Life Science, Pusan National University)
Lee, Hye Ryun (Department of Biomaterials Science, College of Natural Resources & Life Science, Pusan National University)
Choi, Sun Il (Department of Biomaterials Science, College of Natural Resources & Life Science, Pusan National University)
Kwak, Moon Hwa (Department of Biomaterials Science, College of Natural Resources & Life Science, Pusan National University)
Lee, Jae Ho (Department of Biomaterials Science, College of Natural Resources & Life Science, Pusan National University)
Jung, Young Jin (Department of Biomaterials Science, College of Natural Resources & Life Science, Pusan National University)
An, Beum Soo (Department of Biomaterials Science, College of Natural Resources & Life Science, Pusan National University)
Hwang, Dae Youn (Department of Biomaterials Science, College of Natural Resources & Life Science, Pusan National University)
Publication Information
Journal of Life Science / v.23, no.2, 2013 , pp. 167-174 More about this Journal
Abstract
Peroxiredoxin 6 (Prx 6) is a member of the thiol-specific antioxidant protein family, which may play a role in protection against oxidative stress and in regulating phospholipid turnover. The aim of this study was to determine whether a human Prx 6/Luc vector was stably expressed and responded to antioxidants in a lung cell line (NCI-H460). To achieve this, the luciferase signal, hPrx 6 mRNA expression, and superoxide dismutase (SOD) activity were measured in transfectants with a hPrx 6/Luc plasmid after treatment with four antioxidant extracts, including Korea white ginseng (KWG), Korea red ginseng (KRG), Liriope platyphylla (LP), and red Liriope platyphylla (RLP). First, the hPrx 6/Luc plasmid was successfully constructed with DNA fragments of human Prx 6 promoter, amplified by PCR using genomic DNA isolated from NCI-H460 cells, and cloned into the pTransLucent reporter vector. The orientation and sequencing of the hPrx 6/Luc plasmid were identified with restriction enzyme and automatic sequencing. A luciferase assay revealed significant enhancement of luciferase activity in the four treatment groups compared with a vehicle-treated group, although the ratio of the increase was different within each group. The KRG- and LP-treated groups showed higher activity than the KWG- and RLP-treated groups. Furthermore, the luciferase activity against RLP occurred roughly in a dose-dependent manner. However, the level of endogenous hPrx 6 mRNA did not change in any group treated with the four extracts. The SOD activity was in agreement with the luciferase activity. Therefore, these results indicate that the hPrx 6/Luc vector system may successfully express and respond to antioxidant compounds in NCI-H460 cells. The data also suggest that the Prx 6/Luc vector system may be effectively applied in screening the response of hPrx 6 to antioxidant compounds in transgenic mice.
Keywords
Peroxiredoxin 6; luciferase activity; antioxidant; superoxide dismutase (SOD);
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