2024-03-29T05:46:27Zhttp://digital.csic.es/dspace-oai/requestoai:digital.csic.es:10261/777912021-05-24T10:40:40Zcom_10261_64com_10261_1col_10261_317
DIGITAL.CSIC
author
Muñoz-Martínez, F.
author
Lu, P.
author
Cortés-Selva, F.
author
Pérez-Victoria, J. M.
author
Jiménez, I. A.
author
Ravelo, Ángel G.
author
Sharom, F. J.
author
Gamarro, Francisco
author
Castanys, Santiago
2013-06-07T08:52:08Z
2013-06-07T08:52:08Z
2004
Cancer Research 64: 7130- 7138 (2004)
http://hdl.handle.net/10261/77791
10.1158/0008-5472.CAN-04-1005
Overexpression of ABCB1 (MDR1) P-glycoprotein, a multidrug efflux pump, is one mechanism by which tumor cells may develop multidrug resistance (MDR), preventing the successful chemotherapeutic treatment of cancer. Sesquiterpenes from Celastraceae family are natural compounds shown previously to reverse MDR in several human cancer cell lines and Leishmania strains. However, their molecular mechanism of reversion has not been characterized. In the present work, we have studied the ability of 28 dihydro-β-agarofuran Sesquiterpenes to reverse the P-glycoprotein-dependent MDR phenotype and elucidated their molecular mechanism of action. Cytotoxicity assays using human MDR1-transfected NIH-3T3 cells allowed us to select the most potent sesquiterpenes reversing the in vitro resistance to daunomycin and vinblastine. Flow cytometry experiments showed that the above active compounds specifically inhibited drug transport activity of P-glycoprotein in a saturable, concentration-dependent manner (Ki down to 0.24 ± 0.01 μmol/L) but not that of ABCC1 (multidrug resistance protein 1; MRP1), ABCC2 (MRP2), and ABCG2 (breast cancer resistance protein; BCRP) transporters. Moreover, sesquiterpenes inhibited at submicromolar concentrations the P-glycoprotein-mediated transport of [ 3H]colchicine and tetramethylrosamine in plasma membrane from CH RB30 cells and P-glycoprotein-enriched proteoliposomes, supporting that P-glycoprotein is their molecular target. Photoaffinity labeling in plasma membrane and fluorescence spectroscopy experiments with purified protein suggested that sesquiterpenes interact with transmembrane domains of P-glycoprotein. Finally, sesquiterpenes modulated P-glycoprotein ATPase-activity in a biphasic, concentration-dependent manner: they stimulated at very low concentrations but inhibited ATPase activity as noncompetitive inhibitors at higher concentrations. Sesquiterpenes from Celastraceae are promising P-glycoprotein modulators with potential applications in cancer chemotherapy because of their MDR reversal potency and specificity for P-glycoprotein.
eng
closedAccess
Celastraceae sesquiterpenes as a new class of modulators that bind specifically to human P-glycoprotein and reverse cellular multidrug resistance
artículo
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URL
https://digital.csic.es/bitstream/10261/77791/1/accesoRestringido.pdf
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accesoRestringido.pdf
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https://digital.csic.es/bitstream/10261/77791/5/accesoRestringido.pdf.txt
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accesoRestringido.pdf.txt