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Measurements of ion fluxes in breast cell lines using Microelectrode flux estimation

Based on the previous in vitro screening, we identified proprietary crown-ether compound 613 as a potential antitumor drug. Since crown ethers can act as K+ ionophores, we hypothesized that anticancer activity towards breast tumor cells by 613 could be driven by disruption of potassium transport. Potassium homeostasis plays an important role in maintaining of the resting membrane potential, enzyme activities and has an impact on cell proliferation and differentiation. K+channels are involved in the promotion of tumorigenesis. Flow cytometric measurements using the cationic dye DiOC6(3) showed that compound 613 modulates membrane potential in breast cancer stem cell model (HMLE cells). It is extremely difficult to patch clamp HMLE cells, thus we used Microelectrode Flux estimation (MIFE), a non- invasive method based on measuring the concentration changes of ions close to the cells to examine ion fluxes. Specifically, we analysed fluxes of potassium and sodium ions through the plasma membrane of monolayers of HMLE cells and breast tumor cell lines MCF7 and SUM159. Furthermore, membrane potential changes induced by 613 were assessed using fluorescent potentiometric dyes. The results indicate that 613 in fact changes permeability of the plasma membrane of the examined cells for monovalent cations.

Microelectrode flux estimation ; breast cell lines

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