title: DiOC2(3) is not a substrate for multidrug resistance protein (MRP)-mediated drug efflux.
abstract: Multidrug resistance (MDR) is often related to expression of P-glycoprotein (Pgp) or Multidrug Resistance Protein (MRP). Pgp-mediated MDR can be evaluated by determining cellular retention of fluorescent substrates by flow cytometry. This study determined if agents used to evaluate Pgp function also can be used to evaluate MRP function. Cellular retention of doxorubicin (Dox), Rhodamine-123 (Rh-123), and 3,3'-diethyloxacarbocyanine iodide (DiOC2(3)) were studied in MRP-expressing cell lines (HL60/Adr and HT1080/DR4), whereas a Pgp expressing cell line (A2780/Dx5) served as a positive control. Overexpression of Pgp correlated inversely with retention of Dox, Rh-123, and DiOC2(3); however, under identical experimental conditions (1 h reincubation in drug-free medium), no retention difference of the three agents was detected between parental and MRP-expressing resistant cells. Upon extending the reincubation time to 4 h, an efflux of Rh-123 and Dox in the resistant lines became apparent and even more pronounced after 24h; however, still no efflux was detectable for DiOC2(3). Incubation of the cells with a modulator of MDR, PAK-104P, negated the observed drug efflux in Pgp and MRP expressing cells, which correlated with increased sensitivity of the MDR lines to doxorubicin. Thus both Dox and Rh-123 can be used to evaluate MRP-function, but DiOC2(3) can not.
authors.affiliation: Department of Experimental Therapeutics, Grace Cancer Drug Center, Roswell Park Cancer Institute, Buffalo, New York 14263, USA.
authors.forename: H
authors.initials: H
authors.lastname: Minderman
authors.forename: U
authors.initials: U
authors.lastname: Vanhoefer
authors.forename: K
authors.initials: K
authors.lastname: Toth
authors.forename: M B
authors.initials: MB
authors.lastname: Yin
authors.forename: M D
authors.initials: MD
authors.lastname: Minderman
authors.forename: C
authors.initials: C
authors.lastname: Wrzosek
authors.forename: M L
authors.initials: ML
authors.lastname: Slovak
authors.forename: Y M
authors.initials: YM
authors.lastname: Rustum
mesh_terms: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000037:antagonists & inhibitors / Q000378:metabolism*; D001692:Biological Transport; D002232:Carbocyanines / Q000378:metabolism*; D003498:Cyclic P-Oxides / Q000494:pharmacology; D004317:Doxorubicin / Q000378:metabolism; D018432:Drug Resistance, Multiple; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes / Q000378:metabolism*; D006801:Humans; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009539:Nicotinic Acids / Q000494:pharmacology; D020112:Rhodamine 123; D012235:Rhodamines / Q000378:metabolism; D014407:Tumor Cells, Cultured
chemical_list: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D002232:Carbocyanines; D003498:Cyclic P-Oxides; D005456:Fluorescent Dyes; D009363:Neoplasm Proteins; D009539:Nicotinic Acids; D012235:Rhodamines; C103821:PAK 104P; D020112:Rhodamine 123; C081554:3,3'-diethyloxacarbocyanine; D004317:Doxorubicin