Assays for the analysis of P-glycoprotein in acute myeloid leukemia and CD34 subsets of AML blasts.
Expression of the multidrug resistance (MDR) phenotype is an independent prognostic variable in acute myeloid leukemia. Approximately 43-57% of the patients have P-glycoprotein (P-gp) expression. A major drawback with the interpretation of P-gp data in AML is the lack of coherence with different analytical assays. We have focused our efforts of P-gp detection on flow cytometry using a dual technique of P-gp staining with antibodies for the extracellular epitope (MRK16) and a functional analysis of P-gp using the rhodamine efflux assay and the effect of P-gp inhibitors such as SDZ PSC 833. This technique was combined with the staining of lineage-specific antigens such as CD34, CD56 and c-kit. In this way, various subsets of AML cells can be identified such as MRK 16+/-, CD34+/- blasts. These cells can be sorted for further analysis, such as the molecular expression of P-gp and other pleiotropic drug resistance genes.
Sonneveld
P
P
Department of Hematology, University Hospital Rotterdam, The Netherlands.
Wiemer
E
E
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000032:analysis* / Q000235:genetics; D018952:Antigens, CD34 / Q000032:analysis*; D003524:Cyclosporins / Q000494:pharmacology; D005434:Flow Cytometry; D005453:Fluorescence; D006801:Humans; D007150:Immunohistochemistry; D015470:Leukemia, Myeloid, Acute / Q000378:metabolism*; D012313:RNA / Q000032:analysis
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D018952:Antigens, CD34; D003524:Cyclosporins; D012313:RNA; C070272:valspodar