Assessment of multidrug resistance reversal using dielectrophoresis and flow cytometry.
In cancer, multidrug resistance (MDR) is the simultaneous resistance of tumor cells to different natural product anticancer drugs that have no common structure. This is an impediment to the successful treatment of many human cancers. A common correlate of MDR is the overexpression of a membrane protein, P-glycoprotein. Many studies have shown that MDR can be reversed after the use of substrate analogs, called MDR modulators. However, our understanding of MDR modulation is incomplete. In this article, we examine the electrical properties of the human leukemic cells (K562) and its MDR counterpart (K562AR) using dielectrophoresis and flow cytometry (with a membrane potential sensitive dye, DIOC5), both before and after treatment with XR9576 (a P-glycoprotein-specific MDR-reversal agent). The results show significant differences in the cytoplasmic conductivity between the cell lines themselves, but indicate no significant changes after modulation therapy. We conclude that the process of MDR modulation is not associated with changes in the electrical properties of cancer cells. Moreover, the results demonstrate that using the flow cytometry method alone, with MDR cells, may produce artifactual results--whereas in combination with dielectrophoresis, the results show the role of MDR modulators in preventing drug efflux in MDR cells.
Labeed
Fatima H
FH
Centre of Biomedical Engineering, School of Engineering, and Division of Oncology, Postgraduate Medical School, University of Surrey, Guildford, Surrey, United Kingdom.
Coley
Helen M
HM
Thomas
Hilary
H
Hughes
Michael P
MP
D015153:Blotting, Western; D045744:Cell Line, Tumor; D004396:Coloring Agents / Q000494:pharmacology; D003593:Cytoplasm / Q000378:metabolism; D018432:Drug Resistance, Multiple; D019008:Drug Resistance, Neoplasm; D004586:Electrophoresis / Q000379:methods; D005434:Flow Cytometry; D006801:Humans; D020128:Inhibitory Concentration 50; D020014:K562 Cells; D018715:Microscopy, Video; D010641:Phenotype; D011487:Protein Conformation
D004396:Coloring Agents