Stimulation of electroporation-induced inward currents in glioblastoma cell lines by the heat shock protein inhibitor AUY922.

Abstract

Membrane electroporation (MEP) increases the electrical conductivity of the plasma membrane by addition of an external electrical field. Combining MEP-induced current (IMEP ) with antineoplastic agents has been increasingly considered as a new therapeutic manoeuvre, especially in the treatment of malignant gliomas. Thus, the aim of the present study was to evaluate the effect of AUY922 (AUY), a potent inhibitor of heat-shock protein 90 (HSP90), on IMEP in glioblastoma cells. The IMEP in glioblastoma cells (U373) was generated by repetitive hyperpolarization from -80 to -200mV. The amplitude of IMEP was increased by AUY in a concentration-dependent manner, with an EC50 of 0.32μmol/L. In addition AUY shortened the latency to IMEP generation. Before depolarization to +50mV, hyperpolarization to -200mV for 50msec produced Ca(2+) influx and subsequently increased the amplitude of the Ca(2+) -activated K(+) current (IK(Ca) ). The amplitude of IK(Ca) and Ca(2+) influx was further increased by AUY through its ability to activate IMEP . Other HSP90 inhibitors, namely 17-(allylamino)-17-demethoxygeldanamycin (17-AAG; 1μmol/L) and 6-chloro-9-[(4-methoxy-3,5-dimethylpyridin-2-yl)methyl]-9H-purin-2-amine (BIIB021; 1μmol/L), only slightly (albeit significantly) increased the amplitude of IMEP in glioblastoma cells. A 50msec depolarizing step elevated Ca(2+) influx and subsequently increased the amplitude of IK(Ca) in the presence of these three inhibitors. These data indicate that the AUY-mediated stimulation of IMEP and IK(Ca) in glioblastoma cells is independent of HSP90 inhibition. Moreover, these results indicate that AUY-stimulated IMEP and the subsequent activation of IK(Ca) may create important signalling events in glioblastoma cells. Thus, AUY is a drug that could potentially be used to augment the effectiveness of electrochemotherapy.