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

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

Research output: Contribution to journalJournal articleResearchpeer-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
JournalNucleosides Nucleotides & Nucleic Acids
Volume23
Issue number8-9
Pages (from-to)1451-1454
ISSN1525-7770
DOIs
Publication statusPublished - 2004
Externally publishedYes

Keywords

  • cellular efflux
  • breast cancer Breast Neoplasms (MeSH) neoplastic disease, reproductive system disease/female diagnosis, drug therapy, pathology
  • Primates Mammalia Vertebrata Chordata Animalia (Animals, Chordates, Humans, Mammals, Primates, Vertebrates) - Hominidae [86215] 8226/MR20 cell line cell line RPMI 8226/S cell line cell line human myeloid cell
  • ABC-transporter
  • breast cancer resistance protein BCRP
  • Hoechst 33342 23491-52-3
  • Kol43 antineoplastic-drug
  • RPMI
  • 10060, Biochemistry studies - General
  • 12502, Pathology - General
  • 12504, Pathology - Diagnostic
  • 12512, Pathology - Therapy
  • 16506, Reproductive system - Pathology
  • 22002, Pharmacology - General
  • 22005, Pharmacology - Clinical pharmacology
  • 24001, Neoplasms - Diagnostic methods
  • 24004, Neoplasms - Pathology, clinical aspects and systemic effects
  • 24008, Neoplasms - Therapeutic agents and therapy
  • 2-compartment system laboratory techniques
  • Hoechst staining genetic techniques, laboratory techniques
  • Biochemistry and Molecular Biophysics
  • Methods and Techniques
  • Pharmacology

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