| Project/Area Number |
11836007
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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 Institution | Yamaguchi University |
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Principal Investigator |
MUNEKOZU Nakaichi Faculty of Agriculture, Yamaguchi University, Associate Professor, 農学部, 助教授 (60243630)
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| Co-Investigator(Kenkyū-buntansha) |
SATOSHI Une Faculty of Agriculture, Yamaguchi University, Associate Professor, 農学部, 助手 (60294659)
YOSUHO Taura Faculty of Agriculture, Yamaguchi University, Professor, 農学部, 教授 (80163153)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2000: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1999: ¥2,400,000 (Direct Cost: ¥2,400,000)
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| Keywords | dog / lymphoma / antitumor drug / resistance / P-glycoprotein / MDR |
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| Research Abstract |
Canine B cell lymphoma cell line (GL-1) was continuously exposed to the culture medium containing doxorubicin and the cells those could grow even in the presence of the increased level of doxorubicin were obtained (GL-DOXs). In order to characterized the drug resistance observed in GL-DOXs, chemosensitivities to various antineoplastics were investigated with or without verapamil, which reserved P-glycoprotein (P-gp)-mediated drug resistance. The expression of P-gp on the cells were also examined by Western blotting.
GL-DOXs cells were much more resistant not only to doxorubicin and also to vincristine. This resistance was strongly reserved with the presence of verapamil. On the other hand, cisplatin was effective in killing GL-DOXs cells. In the Western blotting. analysis, some protein reacting to the anti-human P-gp monoclonal antibodiy was observed which was absent in GL-1. These results was suggestive enough that GL-DOXs had P-gp on the cell surface and its related gene of MDR (multidrug resistance) was considered to express in the cells.
In the RT-PCR examination using the primers suitable for MDR amplification, a transcript was obtained from the total RNA containing in the GL-DOXs, which was 1281bp with 99% sequence homology to the previously reported canine MDR, 91% to human, 85% to hamster and 80% to mouse and rat. Northern blot analysis using this DNA fragnment revealed a signal at 4200bp, which was identical in size to the cDNA of MDR of human and other spiecies.
GL-DOXs must have multidrug resistance potential mediated by the expression of canine P-gp, and also these cells were considered having canine MDR gene. These cells were considered to be useful models for various investigations on canine multidrug resistance.
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