Effect of phorbol ester on K+ channel in an G292 osteoblast-like cell

Kim, Mi Kyung; Park, Su Byung

Abstract

In order to investigate the action mechanism of protein kinase C on K+ channel in osteoblastic cell, effects of phorbol 12,13-dibutyrate on human osteoblast-like cells (G292) were studied by patch clamp technique with cell-attached configuration. In this experiment, 45pS ion channel was dominant in G292 cell line according to their approximate conductances in symmetrical 140mM KCl saline at holding potential of 60mV. In current-voltage relationship, reversal potential was 5.5mV at the condition of potassium enriched saline in the pipette and -27 mV at the condition of standard extracellular saline in the pipette. Phorbol 12, 13-dibutyrate 10 nM increased the open probability of 45 pS channel and staurosporine, an inhibitor of protein kinase C, suppressed this effect. Phorbol 12,13-dibutyrate moved the reversal potential of 45pS channel to more negative potential and increased the single channel current at the same membrame potential. In order to check the activation of protein kinase C in G292 cell by phorbol 12,13-dibutyrate, western blot of protein kinase C was performed. Phorbol 12,13-dibutyrate 0.1 micrometer translocated protein kinase C from cellular compartment to membrane compartment of the cell. These findings suggest that phorbol 12,13-dibutyrate, one of phorbol esters, activate 45pS channel in G292 cell and affect cell membrane potential, that regulate cellular function.