Saturable function of P-glycoprotein as a drug-efflux pump in multidrug-resistant tumour cells.

P-glycoprotein acts as an active drug-efflux pump in multidrug-resistant tumour cells. We studied the capacity of P-glycoprotein to extrude drugs from the cells. For nanomolar concentrations of vinblastine P388/ADR cells, which overexpress P-glycoprotein in the plasma membrane, accumulated vinblastine, at 37 degrees C for 30 min, to a much lower extent than the sensitive cells (P388/S), while in the micromolar range the cellular concentration was similar for both types of cells. When cells were incubated with a low (10 nM) or high concentration (1 microM) of vinblastine while energy deprived, the vinblastine concentration increased only in the resistant cells incubated with the low concentration of vinblastine, and this increased level was lowered to the level under the normal conditions by addition of glucose. In contrast, the cellular concentrations in other cases were increased to the normal level by glucose. After cells were loaded with the low concentration of vinblastine, the cellular vinblastine was extruded more rapidly from the resistant cells than from the sensitive cells. The courses of vinblastine efflux from the cells loaded with the high concentration of vinblastine were similar in both types of cells. NA-382, a reported P-glycoprotein inhibitor, effectively increased the intracellular vinblastine and inhibited the drug efflux only from multidrug-resistant cells, P388/ADR and AH66 cells, which were incubated with the low concentration of vinblastine. Cellular uptake of NA-382 was also less in P388/ADR cells than in P388/S cells in culture with 10 nM but not 1 microM of the agent, and this low level was reversed to the level in the sensitive cells by 10 microM vinblastine. These results indicate that P-glycoprotein as a drug-efflux pump works effectively under low extracellular concentrations of substrates, but does not under the high concentrations.