| Co-Investigator(Kenkyū-buntansha) |
SAI Yoshimichi Kanazawa University, Graduate School, Assistant Professor, 大学院・自然科学研究科, 助手 (40262589)
TAMAI Ikumi Kanazawa University, Pharmaceutical Sciences, Assistant Professor, 薬学部, 助教授 (20155237)
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| Research Abstract |
In the present study, we studied feasibility of drug delivery using oligopeptide transporter which expressed in some tumor cell lines. During this study period, we obtained following findings.
1. A stable cell line which over expresses human oligopeptide transporter PEPT1 was established by transfecting hPEPTl cDNA into HeLa cells. Its transport actiyity was confirmed by measuring glycylsarcosine and termed the cell line HeLa-hPEPT1.
2. Nude mice (Balb/c nu/nu mice) inoculated with HeLa-hPEPT1 bared tumor within several weeks. In these mice, I.v. injected an anti-tumor drug bestatine, and dipeptide carnosine were accumulated in the hPEPT1-expressing tumors.
3. HeLa-hPEPT1 was more sensitive to bestatine than parent HeLa cell line by in vitro MTT assay. Tiimor growth of HeLa-hPEPT1 in nude mice was strongly suppressed by oral dose of bestatine to the mice, while that of HeLa inoculated mice was not affected.
4. Messenger RNA expression levels of known oligopeptide transporters that is PEPT1 and PEPT2 were quantitatively analyzed by the real time PCR in iridivisual 25 tumbf lines. Among them ML-1, Nakajima and Caco-2 cells expressed PEPT1in high level on the other hand, PEPT2 expression was observed majority of cell lines examined. Comparing mRNA expression and transport activity profiles, we found little correlation between them. Therefore, existence ofnovel transporter that represents the uptake of peptides has been suggested.
5. Molecular cloning of CDNA of organic cation transporter OCTNs and OATPs were performed and some of which were expressed in tumor cell lines.
6. We have proved that expression of amino acid transporter LAT1 and LAT2, which play roles in tumor growth, in the blood-brain barrier.
In conclusion, detailed characterization of many kinds of transporters which expressed in tumors will provide us valuable information to establish method for tumor specific delivery of drugs.
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