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Development and characterization of P-glycoprotein 1 (Pgp1, ABCB1)-mediated doxorubicin-resistant PLHC-1 hepatoma fish cell line

Zaja, Roko, Caminada, Daniel, Lončar, Jovica, Fent, Karl, Smital, Tvrtko·2008·Toxicology and Applied Pharmacology
DOI 10.1016/j.taap.2007.11.001
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Abstract

The development of the multidrug resistance (MDR) phenotype in mammals is often mediated by the overexpression of the P-glycoprotein1 (Pgp, ABCB1) or multidrug resistance-associated protein (MRP)-like ABC transport proteins. A similar phenomenon has also been observed and considered as an important part of the multixenobiotic resistance (MXR) defence system in aquatic organisms. We have recently demonstrated the presence of ABC transporters in the widely used in vitro fish model, the PLHC-1 hepatoma cell line. In the present study we were able to select a highly resistant PLHC-1 sub-clone (PLHC-1/dox) by culturing the wild-type cells in the presence of 1 microM doxorubicin. Using quantitative PCR a 42-fold higher expression of ABCB1 gene was determined in the PLHC-1/dox cells compared to non-selected wild-type cells (PLHC-1/wt). The efflux rates of model fluorescent Pgp1 substrates rhodamine 123 and calcein-AM were 3- to 4-fold higher in the PLHC-1/dox in comparison to the PLHC-1/wt cells. PLHC-1/dox were 45-fold more resistant to doxorubicin cytotoxicity than PLHC-1/wt. Similarly to mammalian cell lines, typical cross-resistance to cytotoxicity of other chemotherapeutics such as daunorubicin, vincristine, vinblastine, etoposide and colchicine, occurred. Furthermore, cyclosporine A, verapamil and PSC833, specific inhibitors of Pgp1 transport activity, completely reversed resistance of PLHC-1/dox cells to all tested drugs, resulting in EC50 values similar to the EC50 values fo…

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Metadata

Authors
Zaja, R., Caminada, D., Lončar, J., Fent, K., & Smital, T.
Venue
Toxicology and Applied Pharmacology
Publisher
Elsevier BV
Published
2008-03
Volume
227
Issue
2
Pages
207-218
ISSN
0041-008X
DOI
10.1016/j.taap.2007.11.001
Cited by
34
References
1
Language
en
Licence
www.elsevier.com/tdm/userlicense/1.0/ (tdm)

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