Open AccessEstablishment of a Stable Cell Line Expressing Green Fluorescence Protein-fused Hypoxia Inducible Factor-1α for Assessment of Carcinogenicity of Chemical Toxicants
Author(s) -
Sung Hoon Kim,
Hiromi Seo,
Minho Lee,
JinHo Chung,
ByungHoon Lee,
MiOck Lee
Publication year - 2009
Publication title -
toxicological research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.872
H-Index - 30
eISSN - 2234-2753
pISSN - 1976-8257
DOI - 10.5487/tr.2009.25.4.189
Subject(s) - green fluorescent protein , cell culture , hypoxia inducible factors , chemistry , carcinogen , hypoxia (environmental) , microbiology and biotechnology , fluorescence , western blot , cancer research , biology , biochemistry , gene , genetics , physics , organic chemistry , quantum mechanics , oxygen
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 CoCl 2 . Well-Known tumor promoters such as mitomycin and methyl methane-sulfonate. 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.