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 language | English (US) |
|---|---|
| Pages (from-to) | 59-65 |
| Number of pages | 7 |
| Journal | Cytometry |
| Volume | 48 |
| Issue number | 2 |
| DOIs | |
| State | Published - Jun 1 2002 |
| Externally published | Yes |
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Keywords
- Breast cancer resistance protein
- Mitoxantrone
- Multidrug resistance
ASJC Scopus subject areas
- Hematology
- Cell Biology
- Pathology and Forensic Medicine
- Biophysics
- Endocrinology
Cite this
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 journal › Article
}
TY - JOUR
T1 - Flow cytometric analysis of breast cancer resistance protein expression and function
AU - Minderman, Hans
AU - Suvannasankha, Attaya
AU - O'Loughlin, Kieran L.
AU - Scheffer, George L.
AU - Scheper, Rik J.
AU - Robey, Robert W.
AU - Baer, Maria R.
PY - 2002/6/1
Y1 - 2002/6/1
N2 - 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.
AB - 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.
KW - Breast cancer resistance protein
KW - Mitoxantrone
KW - Multidrug resistance
UR - http://www.scopus.com/inward/record.url?scp=0036591798&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0036591798&partnerID=8YFLogxK
U2 - 10.1002/cyto.10111
DO - 10.1002/cyto.10111
M3 - Article
VL - 48
SP - 59
EP - 65
JO - Cytometry
JF - Cytometry
SN - 0196-4763
IS - 2
ER -