Possible coexistence of two independent mechanisms contributing to anthracycline resistance in leukaemia P388 cells

Abstract

Murine leukaemia P388 and L1210 cell sublines with varying degrees of resistance to the anthracycline daunomycin (DNM) have been used to monitor (i) intracellular accumulation of DNM, (ii) expression of the drug efflux pump P-glycoprotein (pgp) and (iii) cytoplasmic pH changes. Drug-resistant L1210 65 cells (65-fold resistance), overexpress pgp, and display decreased intracellular accumulation of DNM and identical intracellular pH as compared to the parental drug-sensitive L1210 cell line. On the other hand, moderately drug-resistant P388 20 cells (20-fold resistance), which also exhibit a decreased intracellular drug accumulation with respect to drug-sensitive P388 S cells, display only moderate pgp-encoding mdr1 gene transcription without detectable levels of pgp protein, and undergo cytoplasmic alkalinisation (up to ∼0.2 pH units). A further increase in the level of drug resistance ( P388 100 cells, 100-fold resistance), results in a more pronounced decrease in drug accumulation, significant pgp expression and slightly higher intracellular alkalinisation. Alterations in the degree of protonation of DNM have been shown previously to influence processes such as the rate of uptake and the intracellular accumulation of the drug. On this basis, we propose that the changes in intracellular pH, observed at low levels of drug resistance ( P388 20 cells), could constitute an early cellular response aimed at decreasing the intracellular accumulation of ionisable anti-neoplastics. As the level of resistance increases ( P388 100 ), the cells seem to require more efficient mechanisms of defense against the drug, such as that represented by the expression of pgp. Since there is no apparent correlation between the extent of the changes in intracellular pH and the level of pgp expression in DNM-resistant P388 cell sublines, it is suggested that these two cellular responses contributing to drug resistance could operate independently.