Glucose Derivatives as Efficient Markers of Cell Reversible Electropermeabilization

Molecules spontaneously transported inside the cells, like glucose derivatives, can also be used as electropermeabilization markers. In a previous study, we evaluated the uptake of a fluorescent deoxyglucose derivative (2-NBDG) by normal and electropermeabilized Chinese hamster cells. We extend here our previous study to two murine tumor model cells and investigate the effect of rolipram, a selective PDDE4 inhibitor, on 2-NBDG uptake by tumor cells, with or without electric pulses.2-NBDG was added to cell suspensions, and the cells exposed or not to eight square-wave electric pulses of 100-μs duration and of appropriate field amplitude delivered, were incubated at 37 C and uptake was measured by flow cytometry. In rolipram experiments, cells were similarly processed after a 15 min pre-incubation with rolipram. In spite of significant uptake of 2-NBDG, mediated by GLUT transporters into non permeabilized cells, electric pulses significantly increased the 2-NBDG uptake into both murine tumor cells, even though the electrical parameters allowing a maximal uptake were different. Pretreatment with rolipram, only at high concentrations reduced 2-NBDG uptake in non-electropermeabilized cells, affecting more severely the DC-3F cells than the LPB cells. On the contrary, rolipram treatment did not attenuated the uptake of 2-NBDG in the electropermeabilized cells. We extended to other cell lines our previous observation that glucose derivatives can be used to detect cells reversible electropermeabilization. Moreover, our data suggest that rolipram could probably be used as a tool for improving the visualization of tumor using glucose derivatives, by affecting the uptake in the surrounding normal tissue.