P-glycoprotein-activity measurements in multidrug resistant cell lines

Single-cell versus single-well population fluorescence methods

Jennifer Pasquier, Damien Rioult, Nadine Abu-Kaoud, Sabine Marie, Arash Rafii Tabrizi, Bella S. Guerrouahen, Frank Le Foll

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Background. P-gp expression has been linked to the efflux of chemotherapeutic drugs in human cancers leading to multidrug resistance. Fluorescence techniques have been widely applied to measure the P-gp activity. In this paper, there is a comparison between the advantages of two fluorescence approaches of commonly available and affordable instruments: the microplate reader (MPR) and the flow cytometer to detect the P-gp efflux activity using calcein-AM. Results. The selectivity, sensibility, and reproducibility of the two methods have been defined. Our results showed that the MPR is more powerful for the detection of small inhibition, whereas the flow cytometry method is more reliable at higher concentrations of the inhibitors. We showed that to determine precisely the inhibition efficacy the flow cytometry is better; hence, to get the correct Emax and EC50 values, we cannot only rely on the MPR. Conclusion. Both techniques can potentially be used extensively in the pharmaceutical industry for high-throughput drug screening and in biology laboratories for academic research, monitoring the P-gp efflux in specific assays.

Original languageEnglish
Article number676845
JournalBioMed Research International
Volume2013
DOIs
Publication statusPublished - 2013

Fingerprint

P-Glycoprotein
Flow Cytometry
Flow cytometry
Fluorescence
Cells
Cell Line
Preclinical Drug Evaluations
Drug Industry
Multiple Drug Resistance
Pharmaceutical Preparations
Population
Assays
Screening
Research
Throughput
Neoplasms
Monitoring
Industry
calcein AM

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

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title = "P-glycoprotein-activity measurements in multidrug resistant cell lines: Single-cell versus single-well population fluorescence methods",
abstract = "Background. P-gp expression has been linked to the efflux of chemotherapeutic drugs in human cancers leading to multidrug resistance. Fluorescence techniques have been widely applied to measure the P-gp activity. In this paper, there is a comparison between the advantages of two fluorescence approaches of commonly available and affordable instruments: the microplate reader (MPR) and the flow cytometer to detect the P-gp efflux activity using calcein-AM. Results. The selectivity, sensibility, and reproducibility of the two methods have been defined. Our results showed that the MPR is more powerful for the detection of small inhibition, whereas the flow cytometry method is more reliable at higher concentrations of the inhibitors. We showed that to determine precisely the inhibition efficacy the flow cytometry is better; hence, to get the correct Emax and EC50 values, we cannot only rely on the MPR. Conclusion. Both techniques can potentially be used extensively in the pharmaceutical industry for high-throughput drug screening and in biology laboratories for academic research, monitoring the P-gp efflux in specific assays.",
author = "Jennifer Pasquier and Damien Rioult and Nadine Abu-Kaoud and Sabine Marie and Tabrizi, {Arash Rafii} and Guerrouahen, {Bella S.} and {Le Foll}, Frank",
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journal = "BioMed Research International",
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T1 - P-glycoprotein-activity measurements in multidrug resistant cell lines

T2 - Single-cell versus single-well population fluorescence methods

AU - Pasquier, Jennifer

AU - Rioult, Damien

AU - Abu-Kaoud, Nadine

AU - Marie, Sabine

AU - Tabrizi, Arash Rafii

AU - Guerrouahen, Bella S.

AU - Le Foll, Frank

PY - 2013

Y1 - 2013

N2 - Background. P-gp expression has been linked to the efflux of chemotherapeutic drugs in human cancers leading to multidrug resistance. Fluorescence techniques have been widely applied to measure the P-gp activity. In this paper, there is a comparison between the advantages of two fluorescence approaches of commonly available and affordable instruments: the microplate reader (MPR) and the flow cytometer to detect the P-gp efflux activity using calcein-AM. Results. The selectivity, sensibility, and reproducibility of the two methods have been defined. Our results showed that the MPR is more powerful for the detection of small inhibition, whereas the flow cytometry method is more reliable at higher concentrations of the inhibitors. We showed that to determine precisely the inhibition efficacy the flow cytometry is better; hence, to get the correct Emax and EC50 values, we cannot only rely on the MPR. Conclusion. Both techniques can potentially be used extensively in the pharmaceutical industry for high-throughput drug screening and in biology laboratories for academic research, monitoring the P-gp efflux in specific assays.

AB - Background. P-gp expression has been linked to the efflux of chemotherapeutic drugs in human cancers leading to multidrug resistance. Fluorescence techniques have been widely applied to measure the P-gp activity. In this paper, there is a comparison between the advantages of two fluorescence approaches of commonly available and affordable instruments: the microplate reader (MPR) and the flow cytometer to detect the P-gp efflux activity using calcein-AM. Results. The selectivity, sensibility, and reproducibility of the two methods have been defined. Our results showed that the MPR is more powerful for the detection of small inhibition, whereas the flow cytometry method is more reliable at higher concentrations of the inhibitors. We showed that to determine precisely the inhibition efficacy the flow cytometry is better; hence, to get the correct Emax and EC50 values, we cannot only rely on the MPR. Conclusion. Both techniques can potentially be used extensively in the pharmaceutical industry for high-throughput drug screening and in biology laboratories for academic research, monitoring the P-gp efflux in specific assays.

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