Flow cytometric analysis of breast cancer resistance protein expression and function

Hans Minderman, Attaya Suvannasankha, Kieran L. O'Loughlin, George L. Scheffer, Rik J. Scheper, Robert W. Robey, Maria R. Baer

Research output: Contribution to journalArticle

74 Citations (Scopus)

Abstract

Background: The breast cancer resistance protein (BCRP) is an ATP-binding cassette (ABC) half-transporter that mediates energy-dependent drug efflux. Assessing the clinical relevance of the BCRP will require sensitive and specific methods for detecting its expression and function that allow high-volume specimen throughput and employ widely available instrumentation. Methods: The BXP-34 and BXP-21 monoclonal antibodies were evaluated for flow cytometric detection of BCRP expression. The modulation of efflux of rhodamine-123, 3,3′-diethyloxacarbocyanine iodide, doxorubicin, and mitoxantrone by fumitremorgin C was studied as an assay for BCRP function in BCRP-overexpressing cell lines and controls. Results: BXP-34 and BXP-21 allowed detection of BCRP expression by flow cytometry in all BCRP-expressing cell lines. Mitoxantrone was the only substrate transported by BCRP in all lines, and with mitoxantrone at a 3-μM concentration, light emission (>670 nm) caused by excitation at 488 nm was sufficiently intense to allow detection of differences in retention associated with low levels of BCRP expression. Conclusions: Immunophenotyping with BXP-21 or BXP-34 and fumitremorgin C modulation of mitoxantrone retention allow detection of BCRP expression and function by flow cytometry with standard instrumentation. These assays will facilitate determination of the role of BCRP in clinical drug resistance.

Original languageEnglish (US)
Pages (from-to)59-65
Number of pages7
JournalCytometry
Volume48
Issue number2
DOIs
StatePublished - Jun 1 2002
Externally publishedYes

Fingerprint

Breast Neoplasms
Mitoxantrone
Proteins
Flow Cytometry
Rhodamine 123
Cell Line
Immunophenotyping
ATP-Binding Cassette Transporters
Iodides
Drug Resistance
Doxorubicin
Monoclonal Antibodies
Light
Pharmaceutical Preparations

Keywords

  • Breast cancer resistance protein
  • Mitoxantrone
  • Multidrug resistance

ASJC Scopus subject areas

  • Hematology
  • Cell Biology
  • Pathology and Forensic Medicine
  • Biophysics
  • Endocrinology

Cite this

Minderman, H., Suvannasankha, A., O'Loughlin, K. L., Scheffer, G. L., Scheper, R. J., Robey, R. W., & Baer, M. R. (2002). Flow cytometric analysis of breast cancer resistance protein expression and function. Cytometry, 48(2), 59-65. https://doi.org/10.1002/cyto.10111

Flow cytometric analysis of breast cancer resistance protein expression and function. / Minderman, Hans; Suvannasankha, Attaya; O'Loughlin, Kieran L.; Scheffer, George L.; Scheper, Rik J.; Robey, Robert W.; Baer, Maria R.

In: Cytometry, Vol. 48, No. 2, 01.06.2002, p. 59-65.

Research output: Contribution to journalArticle

Minderman, H, Suvannasankha, A, O'Loughlin, KL, Scheffer, GL, Scheper, RJ, Robey, RW & Baer, MR 2002, 'Flow cytometric analysis of breast cancer resistance protein expression and function', Cytometry, vol. 48, no. 2, pp. 59-65. https://doi.org/10.1002/cyto.10111
Minderman H, Suvannasankha A, O'Loughlin KL, Scheffer GL, Scheper RJ, Robey RW et al. Flow cytometric analysis of breast cancer resistance protein expression and function. Cytometry. 2002 Jun 1;48(2):59-65. https://doi.org/10.1002/cyto.10111
Minderman, Hans ; Suvannasankha, Attaya ; O'Loughlin, Kieran L. ; Scheffer, George L. ; Scheper, Rik J. ; Robey, Robert W. ; Baer, Maria R. / Flow cytometric analysis of breast cancer resistance protein expression and function. In: Cytometry. 2002 ; Vol. 48, No. 2. pp. 59-65.
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