Open AccessDiOC 2 (3) is not a substrate for multidrug resistance protein (MRP)‐mediated drug efflux
Author(s) -
Minderman Hans,
Vanhoefer Udo,
Toth Karoly,
Yin MingBiao,
Minderman M. Denice,
Wrzosek Carol,
Slovak Marilyn L.,
Rustum Youcef M.
Publication year - 1996
Publication title -
cytometry
Language(s) - English
Resource type - Journals
eISSN - 1097-0320
pISSN - 0196-4763
DOI - 10.1002/(sici)1097-0320(19960901)25:1<14::aid-cyto2>3.0.co;2-e
Subject(s) - rhodamine 123 , efflux , multiple drug resistance , p glycoprotein , flow cytometry , doxorubicin , biology , cell culture , multidrug resistance associated proteins , hl60 , drug resistance , microbiology and biotechnology , atp binding cassette transporter , chemistry , biochemistry , in vitro , chemotherapy , transporter , gene , genetics
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 (DiOC 2 (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 DiOC 2 (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 24 h; however, still no efflux was detectable for DiOC 2 (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 DiOC 2 (3) can not. © 1996 Wiley‐Liss, Inc.