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KMID : 0379520090250040189
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한국독성학회지
2009 Volume.25 No. 4 p.189 ~ p.193
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Establishment of a Stable Cell Line Expressing Green Fluorescence Protein-fused Hypoxia Inducible Factor-1α for Assessment of Carcinogenicity of Chemical Toxicants
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Kim Sung-Hye
Seo Hee-Won
Lee Min-Ho
Chung Jin-Ho
Lee Byung-Hoon
Lee Mi-Ock
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Abstract
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Hypoxia inducible factor 1α (HIF-1α) is a potential marker of carcicnogenesis since it is overexpresssed in many human cancers such as brain, breast, and uterus, and its role has implicated in tumor cell growth and metastasis. In this study, we established a stable cell line that express green fluorescence protein (GFP)-fused hypoxia inducible factor 1α (HIF-1α) and evaluated the potential use of this cell line for assessment of carcinogenicity of chemical toxicants. Western blot analysis as well as fluorescence measurements showed that protein-level of GFP-HIF-1α was significantly enhanced in a dose-dependent manner upon treatment of hypoxia mimicking agents such as dexferrioxamine and CoCl₂. Well-Known tumor promoters such as mitomycin and methyl methanesulfonate. significantly induced the fluorescence intensity of GFP-HIF-1α, whereas the known negative controls such as o-anthranilic acid and benzethonium chloride, did not. These results indicate that HIF-1α could be a biological parameter for detection of tumor initiators/promoters and suggest that the GFP-HIF-1α cell line is a useful system for screening of carcinogenic toxicants.
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KEYWORD
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GFP-HIF-1α stable cell line, Chemical toxicants, Carcinogenicity
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