| Project/Area Number |
12671195
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Digestive surgery
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| Research Institution | TOHOKU UNIVERSITY |
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Principal Investigator |
UNNO Michiaki Tohoku University, Graduate School of Medical Science, Research Associate, 大学院・医学系研究科, 助手 (70282043)
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| Co-Investigator(Kenkyū-buntansha) |
ABE Takaaki Tohoku University, Graduate School of Medical Science, Research Associate, 大学院・医学系研究科, 助手 (80292209)
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| Project Period (FY) |
2000 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥4,000,000 (Direct Cost: ¥4,000,000)
Fiscal Year 2001: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2000: ¥3,200,000 (Direct Cost: ¥3,200,000)
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| Keywords | Liver specific organic anion transporter-1 (LST-1) / transcriptional factor / HNF-1a / Luciferase assay / トランスポーター / NF-E2 / HT17 / AP-1 / 遺伝子上流域 / ケノデオキシコール酸 |
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| Research Abstract |
In 1999, we have cloned a novel human organic anion transporter named LST-1. LST-1 is strongly expressed in liver and its substrates are bile acid, conjugated steroids, eicosanoids, and some kinds of drugs. Immunohistochemical analysis indicated that LST-1 was located at sinusoidal membrane of the hepatocytes. The aim of this study is to investigate the transcriptions]-regulation mechanism of human LST-1 gene and its clinical application. The results are follows;
1. Luciferase assay using HT-17 cell lines derived from hepatocyte revealed that AP-1 site (-1100bp upstream from transcriptional starting site) and HNF-1 site (-60bp) were considered to be important region for transcription.
2. Luciferase activity was decreased in the reporter plasmid which inserted point mutation in the HNF-1 binding site.
3. Electrophoresis mobility shift assay (EMSA) using nuclear extract of HT17 revealed that specific protein-DNA complex was detected using oligonucleotide (from-78 to -48).
4. Supershift band was observed by adding a HNF-1 a antibody.
These results indicated that transcriptional factor, HNF-1 a was combined with about -60bp of LST-1 gene upstream and was considered to be most important by transcription of the LST-1 gene in hepatocytes.
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