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Staining with Hoechst 33342 and rhodamine 123: an attempt to detect multidrug resistant phenotype cells in leukemia
S Lahmy1, D Lautier, Y Canitrot
1Université de Perpignan, France.
Abstract:
Development of resistance is the major cause of failure in chemotherapeutic treatments. We have previously shown that the level of labeling with Hoechst 33342 and rhodamine 123 in established cell lines was decreased in cells with 'classic' MDR phenotype. This functional test was carried out using fluorescence image cytometry on living cells. We applied this protocol to patients with chronic lymphocytic leukemia. Although a large variability of the labeling is observed in cells from healthy donors, this approach seems to be useful for early detection of P-gp-dependent resistance in leukemia cells and for identification of new reversing agents on patient lymphocytes.
Insights
Chemotherapy resistance in leukemia can be detected early using a fluorescence imaging test. This method identifies P-glycoprotein-dependent resistance and aids in finding new drugs to reverse it.
Area of Science:
- Oncology
- Pharmacology
- Biotechnology
Background:
- Chemotherapeutic treatment failure is often caused by drug resistance.
- Multidrug resistance (MDR) is a significant challenge in treating cancers, including chronic lymphocytic leukemia (CLL).
- Previous studies indicated reduced labeling with Hoechst 33342 and rhodamine 123 in cell lines with MDR phenotype.
Purpose of the Study:
- To evaluate the utility of a functional fluorescence labeling test for detecting P-glycoprotein (P-gp)-dependent resistance in patient-derived leukemia cells.
- To assess the potential of this method for identifying novel reversing agents for MDR in chronic lymphocytic leukemia.
Main Methods:
- Utilized fluorescence image cytometry on living cells to measure the labeling levels of Hoechst 33342 and rhodamine 123.
- Applied the established protocol to cells from patients with chronic lymphocytic leukemia and compared them to cells from healthy donors.
Main Results:
- Observed significant variability in labeling levels among cells from healthy donors.
- Demonstrated that the fluorescence labeling test can detect P-gp-dependent resistance in leukemia cells.
- The approach showed promise for identifying potential reversing agents using patient lymphocytes.
Conclusions:
- The fluorescence image cytometry method is a valuable tool for the early detection of P-gp-mediated drug resistance in chronic lymphocytic leukemia.
- This functional assay can be applied to patient samples for resistance monitoring and drug discovery efforts.
- Further research into reversing agents is warranted based on these findings.