Determination of multidrug resistance (MDR) activity of
tumor cells could provide important information for the personalized
therapy of
cancer patients. The functional
calcein assay (MultiDrug Quant Assay, Solvo Biotechnology, Budaörs, Hungary) has been proven to be clinically valuable in
hematological malignancies by determining the transporter activity of MDR
protein 1 (MDR1,
ATP-binding cassette
protein [ABC] B1,
P-glycoprotein-170) and MDR-related
protein 1 (
MRP1, ABCC1). In this study, we evaluated if the same functional test was adaptable for the analysis of MDR activity in solid
tumors. For this purpose, tissue specimens of human
colorectal cancer samples were subjected to limited enzymatic digestion by
collagenase to provide a single-cell
suspension; dead cells were excluded by
7-aminoactinomycin D staining, and epithelial
cancer cells were detected by Cy5-conjugated anti-BerEP4
monoclonal antibody. The transporter functions of MDR1 and
MRP1 in viable epithelial cells were assessed by flow cytometry detecting the intracellular accumulation of
calcein dye after exposing cells to various MDR inhibitors.
Collagenase disintegration preserved the MDR activity and the antigenicity of
tumor cells. Thus using the extended
calcein assay provided sufficient viable and functionally active
tumor cells from surgical biopsies to determine the functional MDR activity. In conclusion, the newly described modified
calcein assay may be applicable for evaluating the MDR phenotype in solid tissue specimens from colorectal
forceps biopsy to surgical samples.