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  • Author or Editor: Yoshiko Okai x
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Abstract

Objective—To evaluate the mechanism of multidrug resistance in feline lymphoma cell lines.

Sample Population—A feline lymphoma cell line (FT-1) and its adriamycin (ADM)-resistant subline (FT-1/ADM).

Procedures—The FT-1 cell line was cultivated in the presence of a gradually increasing concentration of ADM to generate its ADM-resistant subline (FT-1/ADM). Susceptibility of cells from the parental FT-1 cell line and the FT-1/ADM subline to antineoplastic drugs was determined. From the complementary DNA (cDNA) template of FT-1/ADM cells, feline MDR1 cDNA was amplified by use of polymerase chain reaction (PCR) and sequenced. Reverse transcription (RT)-PCR and Western blot analyses were performed to assess expression of the MDR1 gene and P-glycoprotein (P-gp) in FT-1/ADM cells, compared with that in FT-1 cells.

Results—A drug sensitivity assay revealed that FT-1/ADM cells were much more resistant to ADM and vincristine than the parental FT-1 cells. The feline MDR1 cDNA amplified by use of PCR was 3,489 base pairs long, corresponding to approximately 90% of the whole open reading frame of human MDR1 cDNA; its amino acid sequence was 91.5, 87.0, and 79.4% identical to that of human MDR1, mouse mdr1a, and mdr1b cDNA, respectively. By RT-PCR analysis, expression of MDR1 messenger RNA was clearly detected in FT-1/ADM cells but not in the parental FT-1 cells. Western blot analysis also revealed the expression of P-gp encoded by the MDR1 gene in FT-1/ADM cells but not in FT-1 cells.

Conclusions—The basic structure of the feline MDR1 gene was essentially the same as that of multidrug- resistance genes of other species. Expression of P-gp appeared to be one of the mechanisms responsible for the development of multidrug resistance in feline lymphoma cell lines in vitro. (Am J Vet Res 2000;61:1122–1127)

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in American Journal of Veterinary Research