2000 Fiscal Year Final Research Report Summary
Proposal and Development of "re-construction artificial liver" using liver-specific organic anion transporter.
| Project/Area Number |
11557088
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Digestive surgery
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| Research Institution | TOHOKU UNIVERSITY |
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Principal Investigator |
MICHIAKI Unno TOHOKU UNIVERSITY, GRADUATE SCHOOL OF MEDICAL SCINECE, ASSOCIATED PROFESSOR (70282043)
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| Co-Investigator(Kenkyū-buntansha) |
ABE Takaaki TOHOKU UNIVERSITY, GRADUATE SCHOOL OF MEDICAL SCINECE, ASSOCIATED PROFESSOR (80292209)
SUZUKI Masanori TOHOKU UNIVERSITY, TOHOKU UNIVERSITY HOSPITAL, ASSOCIATED PROFESSOR (70206530)
ENDO Koujin TOHOKU UNIVERSITY, TOHOKU UNIVERSITY HOSPITAL, ASSOCIATED PROFESSOR (70292315)
KATAYOSE Yu TOHOKU UNIVERSITY, TOHOKU UNIVERSITY HOSPITAL, ASSOCIATED PROFESSOR (20302151)
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| Project Period (FY) |
1999 – 2000
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| Keywords | Liver specific organic anion transporter-1 (LST-1) / LST-2 / hepatocyte / adenovirus / bile acid / artificiall liver / transporter / HepG2 |
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| Research Abstract |
The uptake of various compounds from the circulation is one of the most important functions of hepatocytes. Because of its wide substrate specificity and abundant expression in the liver, human liver-specific organic anion transporter-1 (LST-1) and its subtype LST-2 play important roles in liver function. Both LST-1 and LST-2 are only expressed in liver and its substrates are bile acid, conjugated steroids, eicosanoids, and some kinds of drugs. The aim of this study was to introduce LST-1 and LST-2 recombinant adenovirus into mammalian cells to analyze the function of LST-1 and LST-2 in details. The results are follows;
1. LST-1 and LST-2 cDNA recombinant adenoviruses, termed AdLST-1 and AdLST-2, were constructed by Adenovirus Expression Vector Kit (TaKaRa BIOMEDICALS, Tokyo, Japan).
2. Western blot analysis showed the LST-1 and LST-2 were strongly expressed in HepG2 infected AdLST-1 and AdLST-2. Confocal laser scanning microscopy revealed that LST-1 or LST-2 immunoreactivity was mostly localized at cell membrane, and weakly in the cytosol.
3. Uptake study indicated that LST-1 and LST-2 could transport taurocholate, prostaglandines, methotrexate, dehydroepiandrosterone sulfate, and thyroid hormones. Significant uptake of chenodeoxycholate (CDCA) was detected in LST-2-expressing cell but CDCA transport by LST-1 was not detected.
Our overexpression system using LST-1 or LST-2 recombinant adenovirus can add the liver-specific function of uptaking its specific compounds to the cancer cell line like as HepG2 which has no expression of LST-1 and LST-2.
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