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[Functional detection of P-glycoprotein expressing cells with flow cytometry]
C Ludescher1, W Hilbe, W Eisterer
1Universitätklinik für Innere Medizin, Universität Innsbruck.
Wiener Klinische Wochenschrift
|January 1, 1994
Summary
A novel flow cytometry assay using rhodamine 123 effectively detects multidrug resistance (MDR) in leukemia cells by measuring dye efflux. This functional assay correlates with MDR1 mRNA expression, aiding in cancer treatment strategies.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- Multidrug resistance (MDR) in cancer is often mediated by P-glycoprotein (P-gp) efflux pumps.
- Detecting MDR is crucial for optimizing anti-cancer drug efficacy and patient outcomes.
Purpose of the Study:
- To develop and validate a functional flow cytometry assay for detecting MDR-expressing cells.
- To assess the utility of rhodamine 123 (Rh 123) efflux as a biomarker for MDR in B-cell chronic lymphocytic leukemia (B-CLL) and acute myeloid leukemia (AML).
Main Methods:
- Established a flow cytometry assay using Rh 123 dye to measure drug accumulation and efflux in sensitive and resistant cell lines.
- Analyzed Rh 123 efflux in peripheral blood cells from 42 B-CLL patients and 37 AML patients.
- Correlated Rh 123 efflux assay results with MDR1 mRNA expression levels measured by quantitative polymerase chain reaction (qPCR).
Main Results:
- The Rh 123 assay successfully differentiated drug-sensitive and MDR cells based on dye efflux.
- Significant Rh 123 efflux was observed in 81% of B-CLL patients and 49% of AML patients.
- Rh 123 efflux levels strongly correlated with MDR1 mRNA expression in B-CLL (r = 0.72, p < 0.0001).
Conclusions:
- The rhodamine 123 efflux assay is a sensitive and functional method for detecting MDR in leukemia.
- This assay provides a valuable tool for assessing MDR in clinical settings, potentially guiding therapeutic decisions.