Flow-cytometric determination of intracellular pH, esterase activity and cell volume in human leukemic cell lines following in vitro incubation with cytostatic drugs.
A recently developed flow cytometric assay method using patient tumor cells allows the determination not only of their sensitivity to cytostatic drugs but also of biochemical and biophysical parameters after treatment, such as esterase concentration and intracellular pH of the living cells. DNA-content of the dead cells and cell volume of living and dead cells. The T-cell lines CEM, Molt4, Jurkat, the B-cell lines RPMI1788, Daudi, Raji and the promyelocytic line HL60 were incubated with: cytosine arabinoside (ara-C), L-asparaginase, daunorubicin, vincristine and prednisone for 48 h. Living cells then stained with esterase and pH-dye 1,4-diacetoxy-2,3-dicyanobenzene (ADB) and dead cells with DNA-dye propidium-iodide (PI). The esterase concentration, an index of metabolic activity, decreased in the T-cell lines under the influence of ara-C, daunorubicin and vincristine, whereas in the B-cell lines smaller changes in esterase concentration were observed (P less than 0.001). A decrease in intracellular pH was seen in the ara-C and daunorubicin-incubated cells Molt4, CEM and HL60, whereas in the B-cell lines no significant change in intracellular pH was found. In all lines except Jurkat the cell volume of the surviving cells increased under the influence of certain drugs (primarily ara-C and daunorubicin); B-cell lines showed a greater swelling than T-cell lines (P = 0.001).
Neubauer
A
A
Universitätsklinikum Rudolf-Virchow, Abteilung Innere Medizin, Berlin, FRG.
Valet
G
G
Huhn
D
D
D000970:Antineoplastic Agents / Q000494:pharmacology*; D002470:Cell Survival / Q000187:drug effects; D004351:Drug Resistance; D004354:Drug Screening Assays, Antitumor; D004950:Esterases / Q000032:analysis; D005434:Flow Cytometry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007938:Leukemia / Q000188:drug therapy* / Q000201:enzymology / Q000473:pathology; D014407:Tumor Cells, Cultured
D000970:Antineoplastic Agents; D004950:Esterases