Protein kinase C activity and multidrug resistance in MOLT-3 human lymphoblastic leukemia cells resistant to trimetrexate.
It has been suggested that protein kinase C (PKC) plays a role in multidrug resistance (MDR). In this study we assayed PKC activity in MOLT-3 human acute lymphoblastic leukemia cells and found an approximately 50% decrease in activity in MDR sublines made resistant to the lipophilic antifolate trimetrexate, when compared with trimetrexate-sensitive parent cells. The PKC activity of a methotrexate-resistant subline without MDR (MOLT-3/MTX10,000) was identical to that of parent cells. Although a downward trend was noted in PKC activity in the membrane fraction of cells with increasing trimetrexate resistance, there was no absolute correlation between the degree of MDR and the relative decrease in PKC activity. Using the same method, we also confirmed an over 6-fold increase in PKC activity in the MDR human breast cancer subline MCF-7/DOXR when compared with the sensitive parent cell line, MCF-7/WT. Because of this divergent relationship between relative PKC activity and MDR, we tested the effect of PKC inhibition and activation on drug resistance. The PKC inhibitor staurosporine, at both subtoxic and toxic concentrations as well as at concentrations shown to be inhibitory to PKC, failed to increase drug resistance of parent and resistant MOLT-3 cells and decrease drug resistance of MCF-7/WT and MCF-7/DOXR cells. Short-term exposure to 3-phorbol-12-myristate-13-acetate, which activated PKC 7.0-fold and 4.7-fold, respectively, in the membrane of MOLT-3 and resistant cells, resulted in small (1.3- to 1.8-fold), approximately equivalent, increases (rather than decreases) in resistance to doxorubicin, whereas for vincristine no consistent trend was observed. Identical results were also obtained with phorbol-12,13-dibutyrate. These results indicate that PKC activity can be decreased and increased in MDR cells. Both staurosporine inhibition and phorbol ester activation failed to produce changes in drug resistance that would be considered consistent with the resulting degree of PKC activity. Short-term phorbol ester exposure can change the sensitivity of the cells to doxorubicin without changing the relative drug resistance. PKC activity in these cells may then be unrelated to MDR.
Schwartz
G K
GK
Department of Neoplastic Diseases, Mount Sinai School of Medicine, New York, New York 10029.
Arkin
H
H
Holland
J F
JF
Ohnuma
T
T
D000470:Alkaloids / Q000494:pharmacology; D002455:Cell Division / Q000187:drug effects; D004351:Drug Resistance; D006801:Humans; D066298:In Vitro Techniques; D007945:Leukemia, Lymphoid / Q000201:enzymology / Q000473:pathology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011799:Quinazolines / Q000494:pharmacology*; D019311:Staurosporine; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D016597:Trimetrexate; D014407:Tumor Cells, Cultured / Q000187:drug effects* / Q000201:enzymology
D000470:Alkaloids; D011799:Quinazolines; D011493:Protein Kinase C; D019311:Staurosporine; D013755:Tetradecanoylphorbol Acetate; D016597:Trimetrexate