HIF-1-Dependent VEGF Reporter Gene Assay by a Stable Transformant of CHO Cells
スポンサーリンク
概要
- 論文の詳細を見る
For the establishment of a screening system to detect inhibitors of vascular endotherial growth factor (VEGF) expression, a stable transformant of Chinese hamster ovary cells was isolated and cloned by transaction of a hypoxia-inducible factor 1 (HIF-1)-dependent VEGF promoter reporter gene. The expression of the reporter gene in the clone cells, as measured by luciferase activity, was stable. Hypoxic responses were best observed at an initial cell density of 2 × 10^4/well. The maximal increase of luciferase activity was 30 fold. In the highest cell density of 8 × 10^4/well (2.1 × 10^5/cm^2), basal activity was increased 13-15 fold compared to that at the lower cell densities, and did not respond to hypoxia. Addition of CoCl_2, which is known to mimic hypoxia, increased luciferase activity more than 10 times in normoxia. Nitric oxide donors, which arc known to suppress the activation of HIF-1, inhibited expression of the VEGF promoter reporter gene under hypoxia. Histone deacetylase inhibitors, trichostatin A and sodium ≪-butyrate which are known to stimulate transcription of many genes enhanced its transcription in hypoxia. These results indicate that the stable transformant is a useful tool for screening of HIF-1 modifiers.
- 公益社団法人日本薬学会の論文
- 2003-04-01
著者
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TAKEUCHI Tomio
Institute of Microbial Chemistry
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Kondo S
Institute Of Microbial Chemistry
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Takeuchi T
Bioresource Laboratories Mercian Corp.
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Takeuchi T
Institute Of Microbial Chemistry
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KUNIMOTO Setsuko
Institute of Microbial Chemistry
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EGAWA Kiyoshi
Department of Microbiology, Showa University of Pharmaceutical Sciences
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Yamazaki Yohko
微生物化学研究会微生物化学研究センター沼津創薬医科学研究所
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Nose K
Department Of Microbiology Showa University Of Pharmaceutical Sciences
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Nose K
Showa Univ. Pharmaceutical Sci. Tokyo Jpn
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KUNIMOTO Setsuko
Numazu Bio-Medical Research Institute, Microbial Chemistry Research Center
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YAMAZAKI Yhko
Institute of Microbial Chemistry
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EGAWA Kiyoshi
Showa University, School of Pharmaceutical Science
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NOSE Kiyoshi
Showa University, School of Pharmaceutical Science
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TAKIZAWA Toshio
Teikyo University Institute of Medical Mycology
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Takeuchi Toshio
Research And Development Department Kibun Food Chemifa Co. Ltd.
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Nose Kiyoshi
Department Of Microbiology Showa University Of Pharmaceutical Sciences
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Yamazaki Yoko
Numazu Bio-medical Research Institute Microbial Chemistry Research Center
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Egawa K
Department Of Microbiology Showa University Of Pharmaceutical Sciences
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YAMAZAKI Yohko
Institute of Microbial Chemistry
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Kunimoto S
Numazu Bio-medical Research Institute Microbial Chemistry Research Center
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山嵜 行由
School Of Pharmaceutical Sciences University Of Shizuoka
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Takeuchi Tomio
Institute For Chemotherapy M.c.r.f.
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Nose Kiyoshi
Showa Univ. School Of Pharmacy Tokyo
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Takeuchi T
Inst. Microbial Chemistry Shinagawa‐ku Jpn
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