Biophysical characterization of MDR breast cancer cell lines reveals the cytoplasm is critical in determining drug sensitivity

Abstract

Dielectrophoresis (DEP) was used to examine a panel of MCF-7 cell lines comprising parental MCF-7 cells and MDR derivatives: MCF-7TaxR (paclitaxel-resistant, P-glycoprotein (P-gp) positive), MCF-7DoxR (doxorubicin-resistant MRP2 positive) plus MCF-7 MDR1 ( MDR1 transfected, P-gp positive). MCF-7DoxR and MCF-7 MDR1 were broadly cross-resistant to natural product anticancer agents, whereas MCF-7TaxR cells were not, contrary to P-gp expression. Whilst DEP revealed modest membrane changes in MDR sub-lines, we saw significant changes in their cytoplasmic conductivity: MCF-7TaxR < MCF-7 < MCF-7MDR1 < MCF-7DoxR (range 0.14–0.40 S/m). Cytoplasmic conductivity is affected by the movement of molecules e.g. as in intracellular trafficking MCF-7TaxR showed a reduced membrane potential, whereas MCF-7DoxR and MCF-7 MDR1 showed an increase. Thus, altered membrane potential is associated with an MDR phenotype, but in a complex manner. DEP data suggest a model whereby relative increases in cytoplasmic conductivity are correlated with MDR, whilst relative decreases equate with a sensitised phenotype e.g. MCF-7TaxR. Moreover, extent of anthracycline accumulation was inversely related to cytoplasmic conductivity. These data are representative of a model where drug sensitivity is associated with low ionic conductance (reduced cellular trafficking and ion transport) and substantial anthracycline accumulation. For classical MDR i.e. MCF-7 MDR1, we saw the reverse picture. Thus, the drug resistance phenotypes of this panel of MCF-7 lines can be delineated by assessment of cytoplasmic biophysical properties using DEP.