Online fluorescent method to assess BCRP/ABCG2 activity in suspension cells

J H Hooijberg, G J Peters, G J L Kaspers, P R Wielinga, A J P Veerman, R Pieters, G Jansen

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

An online method was developed to monitor BCRP mediated efflux of fluorescent substrates in suspension cells. To this end, a 2-compartment system consisting of a transwell cup and a cuvette was used. In this system we were able to observe differences in efflux kinetics between BCRP overexpressing RPMI 8226/MR cells and parental myeloid RPMI 8226(s) cells using only 50,000 cells per experiment. 8226/MR cells displayed a larger cellular efflux rate of the BCRP substrate Hoechst 33342, as compared to the wildtype cells. This difference in efflux rate was completely decreased in the presence of the BCRP inhibitor Ko143.

Original languageEnglish
Pages (from-to)1451-4
Number of pages4
JournalNucleosides, Nucleotides and Nucleic Acids
Volume23
Issue number8-9
DOIs
Publication statusPublished - Oct 2004

Cite this

@article{96818f4c600d4bdc8911e28d3d0c2d78,
title = "Online fluorescent method to assess BCRP/ABCG2 activity in suspension cells",
abstract = "An online method was developed to monitor BCRP mediated efflux of fluorescent substrates in suspension cells. To this end, a 2-compartment system consisting of a transwell cup and a cuvette was used. In this system we were able to observe differences in efflux kinetics between BCRP overexpressing RPMI 8226/MR cells and parental myeloid RPMI 8226(s) cells using only 50,000 cells per experiment. 8226/MR cells displayed a larger cellular efflux rate of the BCRP substrate Hoechst 33342, as compared to the wildtype cells. This difference in efflux rate was completely decreased in the presence of the BCRP inhibitor Ko143.",
keywords = "ATP Binding Cassette Transporter, Subfamily G, Member 2, ATP-Binding Cassette Transporters/chemistry, Benzimidazoles/pharmacology, Biological Transport, Cell Line, Tumor, Cells, Cultured, Fluorescent Dyes/pharmacology, Humans, Kinetics, Neoplasm Proteins/chemistry, Radiation-Sensitizing Agents/pharmacology, Software, Spectrometry, Fluorescence/methods, Time Factors",
author = "Hooijberg, {J H} and Peters, {G J} and Kaspers, {G J L} and Wielinga, {P R} and Veerman, {A J P} and R Pieters and G Jansen",
year = "2004",
month = "10",
doi = "10.1081/NCN-200027672",
language = "English",
volume = "23",
pages = "1451--4",
journal = "Nucleosides, Nucleotides and Nucleic Acids",
issn = "1525-7770",
publisher = "Taylor and Francis Ltd.",
number = "8-9",

}

Online fluorescent method to assess BCRP/ABCG2 activity in suspension cells. / Hooijberg, J H; Peters, G J; Kaspers, G J L; Wielinga, P R; Veerman, A J P; Pieters, R; Jansen, G.

In: Nucleosides, Nucleotides and Nucleic Acids, Vol. 23, No. 8-9, 10.2004, p. 1451-4.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Online fluorescent method to assess BCRP/ABCG2 activity in suspension cells

AU - Hooijberg, J H

AU - Peters, G J

AU - Kaspers, G J L

AU - Wielinga, P R

AU - Veerman, A J P

AU - Pieters, R

AU - Jansen, G

PY - 2004/10

Y1 - 2004/10

N2 - An online method was developed to monitor BCRP mediated efflux of fluorescent substrates in suspension cells. To this end, a 2-compartment system consisting of a transwell cup and a cuvette was used. In this system we were able to observe differences in efflux kinetics between BCRP overexpressing RPMI 8226/MR cells and parental myeloid RPMI 8226(s) cells using only 50,000 cells per experiment. 8226/MR cells displayed a larger cellular efflux rate of the BCRP substrate Hoechst 33342, as compared to the wildtype cells. This difference in efflux rate was completely decreased in the presence of the BCRP inhibitor Ko143.

AB - An online method was developed to monitor BCRP mediated efflux of fluorescent substrates in suspension cells. To this end, a 2-compartment system consisting of a transwell cup and a cuvette was used. In this system we were able to observe differences in efflux kinetics between BCRP overexpressing RPMI 8226/MR cells and parental myeloid RPMI 8226(s) cells using only 50,000 cells per experiment. 8226/MR cells displayed a larger cellular efflux rate of the BCRP substrate Hoechst 33342, as compared to the wildtype cells. This difference in efflux rate was completely decreased in the presence of the BCRP inhibitor Ko143.

KW - ATP Binding Cassette Transporter, Subfamily G, Member 2

KW - ATP-Binding Cassette Transporters/chemistry

KW - Benzimidazoles/pharmacology

KW - Biological Transport

KW - Cell Line, Tumor

KW - Cells, Cultured

KW - Fluorescent Dyes/pharmacology

KW - Humans

KW - Kinetics

KW - Neoplasm Proteins/chemistry

KW - Radiation-Sensitizing Agents/pharmacology

KW - Software

KW - Spectrometry, Fluorescence/methods

KW - Time Factors

U2 - 10.1081/NCN-200027672

DO - 10.1081/NCN-200027672

M3 - Article

VL - 23

SP - 1451

EP - 1454

JO - Nucleosides, Nucleotides and Nucleic Acids

JF - Nucleosides, Nucleotides and Nucleic Acids

SN - 1525-7770

IS - 8-9

ER -