Accurate measurement of
P-glycoprotein (P-170) expression in clinical samples still remains a controversial issue. In this study
tumor cell P-170 expression was assessed in 29 patients suffering from acute
leukemia (17
acute myeloid leukemia (AML) and 12
acute lymphoblastic leukemia (ALL)) using three different techniques: flow cytometry measuring
rhodamine 123 (Rh123) efflux (functional level), immunocytochemistry (
protein level) and RT-PCR (
mRNA level). Rh123 efflux was detectable in 10/29 (34%) of all cases, in 9/17 (53%) of AML and in 1/12 (8%) of ALL samples. In AML patients a significant association of CD34 expression and P-170 activity was observed (P < 0.02). All AML patients with the FAB subtype M5 were Rh123 negative (P < 0.007). Cytospin preparations were analyzed for staining with
monoclonal antibodies JSB1 and MM4.17. Eight of 16 (50%) AML and 0/9 (0%) ALL cases expressed the multidrug resistance (MDR)
protein assessed by JSB1. With MM4.17 87% of AML and 50% of ALL patients were scored positive. Agreement between both
antibodies was found in only 13/23 (57%) samples. Extracted
RNA from 12 patients was analyzed by RT-PCR to evaluate the expression of MDR1 and
multidrug resistance-associated protein (MRP)
mRNA. An increased level of MDR1
mRNA was detectable in 4/7 AML and 0/5 ALL cases. MRP expression was found in 3/7 AML and 0/5 ALL patients. Comparison of Rh123 assay and immunocytochemistry revealed a very good correlation when using MoAb JSB1 (P < 0.004) but not with MM4.17 (not significant (NS)). JSB1 also showed a much better association with the PCR results (P < 0.05) than MM4.17 (NS). Finally, we compared the results of the functional Rh123 assay and RT-PCR and observed a high correlation for Rh123/MDR1 (r = 0.819, P < 0.001) but low for Rh123/MRP (r = 0.562, NS). We conclude that measurement of Rh123 efflux and immunocytochemical staining of cytospin preparations with JSB1 allows the accurate monitoring of P-170 expression in acute
leukemia. The simplicity of these two MDR assays suggests their use for routine MDR screening.