Demonstration of the activity of P-glycoprotein by a semi-automated fluorometric method

G. Spengler, Miguel Viveiros, Marta Martins, Liliana Rodrigues, Ana Martins, J. Molnár, Isabel Couto, Leonard Amaral

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Background: We have developed a semi-automated fluorometric method that utilizes ethidium bromide (EB), a common substrate of bacterial efflux pumps. The method is sufficiently sensitive to characterize the efflux pump systems of bacteria. Because EB is also recognized and extruded by ATP-binding cassette (ABC) transporters and these have similarity to P-glycoprotein (P-gp), the method has been extended for the evaluation of agents that can inhibit the extrusion of EB on a real-time basis by mouse lymphoma cells containing the human ABCB1 (mdr1) gene. Materials and Methods: Monitoring of uptake and extrusion of EB was assessed using the Rotor-Gene™ 3000 (Corbett Research) under different conditions. Results: Whereas extrusion of EB took place readily, the addition of known inhibitors of efflux pumps (verapamil, reserpine) caused retention of EB. Conclusion: This method is inexpensive and allows the detection of neoplastic cells with increased efflux activity as well as the screening of large numbers of compounds for inhibition of the P-gp.

Original languageEnglish
Pages (from-to)2173-2177
Number of pages5
JournalAnticancer Research
Volume29
Issue number6
Publication statusPublished - Jun 2009

Fingerprint

Ethidium
P-Glycoprotein
ATP-Binding Cassette Transporters
Reserpine
Verapamil
Genes
Lymphoma
Bacteria
Research

Keywords

  • ABC transporters
  • ABCB1
  • Efflux
  • Ethidium bromide
  • mdr1
  • Mouse lymphoma cells
  • P-glycoprotein
  • Semi-automated fluorometric method

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Spengler, G., Viveiros, M., Martins, M., Rodrigues, L., Martins, A., Molnár, J., ... Amaral, L. (2009). Demonstration of the activity of P-glycoprotein by a semi-automated fluorometric method. Anticancer Research, 29(6), 2173-2177.

Demonstration of the activity of P-glycoprotein by a semi-automated fluorometric method. / Spengler, G.; Viveiros, Miguel; Martins, Marta; Rodrigues, Liliana; Martins, Ana; Molnár, J.; Couto, Isabel; Amaral, Leonard.

In: Anticancer Research, Vol. 29, No. 6, 06.2009, p. 2173-2177.

Research output: Contribution to journalArticle

Spengler, G, Viveiros, M, Martins, M, Rodrigues, L, Martins, A, Molnár, J, Couto, I & Amaral, L 2009, 'Demonstration of the activity of P-glycoprotein by a semi-automated fluorometric method', Anticancer Research, vol. 29, no. 6, pp. 2173-2177.
Spengler, G. ; Viveiros, Miguel ; Martins, Marta ; Rodrigues, Liliana ; Martins, Ana ; Molnár, J. ; Couto, Isabel ; Amaral, Leonard. / Demonstration of the activity of P-glycoprotein by a semi-automated fluorometric method. In: Anticancer Research. 2009 ; Vol. 29, No. 6. pp. 2173-2177.
@article{25c2a95a5a084ff0a75ff3f1aaa29243,
title = "Demonstration of the activity of P-glycoprotein by a semi-automated fluorometric method",
abstract = "Background: We have developed a semi-automated fluorometric method that utilizes ethidium bromide (EB), a common substrate of bacterial efflux pumps. The method is sufficiently sensitive to characterize the efflux pump systems of bacteria. Because EB is also recognized and extruded by ATP-binding cassette (ABC) transporters and these have similarity to P-glycoprotein (P-gp), the method has been extended for the evaluation of agents that can inhibit the extrusion of EB on a real-time basis by mouse lymphoma cells containing the human ABCB1 (mdr1) gene. Materials and Methods: Monitoring of uptake and extrusion of EB was assessed using the Rotor-Gene™ 3000 (Corbett Research) under different conditions. Results: Whereas extrusion of EB took place readily, the addition of known inhibitors of efflux pumps (verapamil, reserpine) caused retention of EB. Conclusion: This method is inexpensive and allows the detection of neoplastic cells with increased efflux activity as well as the screening of large numbers of compounds for inhibition of the P-gp.",
keywords = "ABC transporters, ABCB1, Efflux, Ethidium bromide, mdr1, Mouse lymphoma cells, P-glycoprotein, Semi-automated fluorometric method",
author = "G. Spengler and Miguel Viveiros and Marta Martins and Liliana Rodrigues and Ana Martins and J. Moln{\'a}r and Isabel Couto and Leonard Amaral",
year = "2009",
month = "6",
language = "English",
volume = "29",
pages = "2173--2177",
journal = "Anticancer Research",
issn = "0250-7005",
publisher = "International Institute of Anticancer Research",
number = "6",

}

TY - JOUR

T1 - Demonstration of the activity of P-glycoprotein by a semi-automated fluorometric method

AU - Spengler, G.

AU - Viveiros, Miguel

AU - Martins, Marta

AU - Rodrigues, Liliana

AU - Martins, Ana

AU - Molnár, J.

AU - Couto, Isabel

AU - Amaral, Leonard

PY - 2009/6

Y1 - 2009/6

N2 - Background: We have developed a semi-automated fluorometric method that utilizes ethidium bromide (EB), a common substrate of bacterial efflux pumps. The method is sufficiently sensitive to characterize the efflux pump systems of bacteria. Because EB is also recognized and extruded by ATP-binding cassette (ABC) transporters and these have similarity to P-glycoprotein (P-gp), the method has been extended for the evaluation of agents that can inhibit the extrusion of EB on a real-time basis by mouse lymphoma cells containing the human ABCB1 (mdr1) gene. Materials and Methods: Monitoring of uptake and extrusion of EB was assessed using the Rotor-Gene™ 3000 (Corbett Research) under different conditions. Results: Whereas extrusion of EB took place readily, the addition of known inhibitors of efflux pumps (verapamil, reserpine) caused retention of EB. Conclusion: This method is inexpensive and allows the detection of neoplastic cells with increased efflux activity as well as the screening of large numbers of compounds for inhibition of the P-gp.

AB - Background: We have developed a semi-automated fluorometric method that utilizes ethidium bromide (EB), a common substrate of bacterial efflux pumps. The method is sufficiently sensitive to characterize the efflux pump systems of bacteria. Because EB is also recognized and extruded by ATP-binding cassette (ABC) transporters and these have similarity to P-glycoprotein (P-gp), the method has been extended for the evaluation of agents that can inhibit the extrusion of EB on a real-time basis by mouse lymphoma cells containing the human ABCB1 (mdr1) gene. Materials and Methods: Monitoring of uptake and extrusion of EB was assessed using the Rotor-Gene™ 3000 (Corbett Research) under different conditions. Results: Whereas extrusion of EB took place readily, the addition of known inhibitors of efflux pumps (verapamil, reserpine) caused retention of EB. Conclusion: This method is inexpensive and allows the detection of neoplastic cells with increased efflux activity as well as the screening of large numbers of compounds for inhibition of the P-gp.

KW - ABC transporters

KW - ABCB1

KW - Efflux

KW - Ethidium bromide

KW - mdr1

KW - Mouse lymphoma cells

KW - P-glycoprotein

KW - Semi-automated fluorometric method

UR - http://www.scopus.com/inward/record.url?scp=67650895867&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=67650895867&partnerID=8YFLogxK

M3 - Article

C2 - 19528478

AN - SCOPUS:67650895867

VL - 29

SP - 2173

EP - 2177

JO - Anticancer Research

JF - Anticancer Research

SN - 0250-7005

IS - 6

ER -