Characterization of functional assays of multidrug resistance P-glycoprotein transport activity

I. Bosch, C. L. Crankshaw, D. Piwnica-Worms, James Croop

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

P-glycoprotein-mediated multidrug resistance has emerged as one of the most attractive targets to improve anticancer therapy. The P-glycoprotein functions as an energy-dependent, membrane transport pump capable of decreasing the intracellular concentration of a broad range of chemotherapeutic agents. Pharmaceuticals which inhibit P-glycoprotein transport activity are currently being evaluated in clinical trials. Characterization of P-glycoprotein functional activity is critical in determining if these multidrug resistance reversal agents improve therapeutic responses of tumors expressing P-glycoprotein. In this report, we directly compare and characterize assays using rhodamine 123, dimethyloxadicarbocyanine iodide (DiOC2), [3H]daunorubicin and hexakis(2-methoxyisobutyl isonitrile)technetium(I) ([99mTc]Sestamibi) as P-glycoprotein transport probes to quantitate functional activity. The accumulation of certain substrates is concentration dependent and the parameters which determine probe accumulation are impacted by the level of P-glycoprotein expression. In addition, higher concentrations of reversal agents are required to inhibit multidrug resistance in cell lines expressing higher levels of P-glycoprotein. Furthermore, the concentration of reversal agents required to inhibit completely P-glycoprotein transport activity is higher than generally recognized. Thus, the level of P-glycoprotein expression may confound intersample comparisons unless sensitive probes are used in combination with saturating concentrations of potent reversal agents. These results highlight the importance of carefully characterizing assay systems under uniform conditions to quantitate P-glycoprotein function.

Original languageEnglish (US)
Pages (from-to)1131-1137
Number of pages7
JournalLeukemia
Volume11
Issue number7
StatePublished - 1997
Externally publishedYes

Fingerprint

Multiple Drug Resistance
P-Glycoprotein
Rhodamine 123
Technetium Tc 99m Sestamibi
Daunorubicin
Technetium
Iodides
Clinical Trials
Cell Line

Keywords

  • MDR
  • Multidrug resistance
  • P-glycoprotein

ASJC Scopus subject areas

  • Hematology
  • Cancer Research

Cite this

Characterization of functional assays of multidrug resistance P-glycoprotein transport activity. / Bosch, I.; Crankshaw, C. L.; Piwnica-Worms, D.; Croop, James.

In: Leukemia, Vol. 11, No. 7, 1997, p. 1131-1137.

Research output: Contribution to journalArticle

Bosch, I, Crankshaw, CL, Piwnica-Worms, D & Croop, J 1997, 'Characterization of functional assays of multidrug resistance P-glycoprotein transport activity', Leukemia, vol. 11, no. 7, pp. 1131-1137.
Bosch, I. ; Crankshaw, C. L. ; Piwnica-Worms, D. ; Croop, James. / Characterization of functional assays of multidrug resistance P-glycoprotein transport activity. In: Leukemia. 1997 ; Vol. 11, No. 7. pp. 1131-1137.
@article{c62d0e109eac44c18c30831f9bac9255,
title = "Characterization of functional assays of multidrug resistance P-glycoprotein transport activity",
abstract = "P-glycoprotein-mediated multidrug resistance has emerged as one of the most attractive targets to improve anticancer therapy. The P-glycoprotein functions as an energy-dependent, membrane transport pump capable of decreasing the intracellular concentration of a broad range of chemotherapeutic agents. Pharmaceuticals which inhibit P-glycoprotein transport activity are currently being evaluated in clinical trials. Characterization of P-glycoprotein functional activity is critical in determining if these multidrug resistance reversal agents improve therapeutic responses of tumors expressing P-glycoprotein. In this report, we directly compare and characterize assays using rhodamine 123, dimethyloxadicarbocyanine iodide (DiOC2), [3H]daunorubicin and hexakis(2-methoxyisobutyl isonitrile)technetium(I) ([99mTc]Sestamibi) as P-glycoprotein transport probes to quantitate functional activity. The accumulation of certain substrates is concentration dependent and the parameters which determine probe accumulation are impacted by the level of P-glycoprotein expression. In addition, higher concentrations of reversal agents are required to inhibit multidrug resistance in cell lines expressing higher levels of P-glycoprotein. Furthermore, the concentration of reversal agents required to inhibit completely P-glycoprotein transport activity is higher than generally recognized. Thus, the level of P-glycoprotein expression may confound intersample comparisons unless sensitive probes are used in combination with saturating concentrations of potent reversal agents. These results highlight the importance of carefully characterizing assay systems under uniform conditions to quantitate P-glycoprotein function.",
keywords = "MDR, Multidrug resistance, P-glycoprotein",
author = "I. Bosch and Crankshaw, {C. L.} and D. Piwnica-Worms and James Croop",
year = "1997",
language = "English (US)",
volume = "11",
pages = "1131--1137",
journal = "Leukemia",
issn = "0887-6924",
publisher = "Nature Publishing Group",
number = "7",

}

TY - JOUR

T1 - Characterization of functional assays of multidrug resistance P-glycoprotein transport activity

AU - Bosch, I.

AU - Crankshaw, C. L.

AU - Piwnica-Worms, D.

AU - Croop, James

PY - 1997

Y1 - 1997

N2 - P-glycoprotein-mediated multidrug resistance has emerged as one of the most attractive targets to improve anticancer therapy. The P-glycoprotein functions as an energy-dependent, membrane transport pump capable of decreasing the intracellular concentration of a broad range of chemotherapeutic agents. Pharmaceuticals which inhibit P-glycoprotein transport activity are currently being evaluated in clinical trials. Characterization of P-glycoprotein functional activity is critical in determining if these multidrug resistance reversal agents improve therapeutic responses of tumors expressing P-glycoprotein. In this report, we directly compare and characterize assays using rhodamine 123, dimethyloxadicarbocyanine iodide (DiOC2), [3H]daunorubicin and hexakis(2-methoxyisobutyl isonitrile)technetium(I) ([99mTc]Sestamibi) as P-glycoprotein transport probes to quantitate functional activity. The accumulation of certain substrates is concentration dependent and the parameters which determine probe accumulation are impacted by the level of P-glycoprotein expression. In addition, higher concentrations of reversal agents are required to inhibit multidrug resistance in cell lines expressing higher levels of P-glycoprotein. Furthermore, the concentration of reversal agents required to inhibit completely P-glycoprotein transport activity is higher than generally recognized. Thus, the level of P-glycoprotein expression may confound intersample comparisons unless sensitive probes are used in combination with saturating concentrations of potent reversal agents. These results highlight the importance of carefully characterizing assay systems under uniform conditions to quantitate P-glycoprotein function.

AB - P-glycoprotein-mediated multidrug resistance has emerged as one of the most attractive targets to improve anticancer therapy. The P-glycoprotein functions as an energy-dependent, membrane transport pump capable of decreasing the intracellular concentration of a broad range of chemotherapeutic agents. Pharmaceuticals which inhibit P-glycoprotein transport activity are currently being evaluated in clinical trials. Characterization of P-glycoprotein functional activity is critical in determining if these multidrug resistance reversal agents improve therapeutic responses of tumors expressing P-glycoprotein. In this report, we directly compare and characterize assays using rhodamine 123, dimethyloxadicarbocyanine iodide (DiOC2), [3H]daunorubicin and hexakis(2-methoxyisobutyl isonitrile)technetium(I) ([99mTc]Sestamibi) as P-glycoprotein transport probes to quantitate functional activity. The accumulation of certain substrates is concentration dependent and the parameters which determine probe accumulation are impacted by the level of P-glycoprotein expression. In addition, higher concentrations of reversal agents are required to inhibit multidrug resistance in cell lines expressing higher levels of P-glycoprotein. Furthermore, the concentration of reversal agents required to inhibit completely P-glycoprotein transport activity is higher than generally recognized. Thus, the level of P-glycoprotein expression may confound intersample comparisons unless sensitive probes are used in combination with saturating concentrations of potent reversal agents. These results highlight the importance of carefully characterizing assay systems under uniform conditions to quantitate P-glycoprotein function.

KW - MDR

KW - Multidrug resistance

KW - P-glycoprotein

UR - http://www.scopus.com/inward/record.url?scp=0030795841&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0030795841&partnerID=8YFLogxK

M3 - Article

C2 - 9205002

AN - SCOPUS:0030795841

VL - 11

SP - 1131

EP - 1137

JO - Leukemia

JF - Leukemia

SN - 0887-6924

IS - 7

ER -