Isolation and Characterization of an Anthracycline-resistant Human Leukemic Cell Line1
نویسندگان
چکیده
An anthracycline-resistant subline of HL-60 promyelocytic leu kemia cells (HL-60/AR) has been isolated in vitro by subculturing in progressively higher concentrations of Adriamycin. The resis tant cells are capable of sustaining continuous growth in 10~6 M Adriamycin which is more than 50 times the 50% inhibitory dose for the parent line. HL-60/AR expressed variable degrees of cross-resistance to daunorubicin, dihydroxyanthracenedione, vincristine, vinblastine, and actinomycin D, but it remained sen sitive to methotrexate and 1-0-D-arabinofuranosylcytosine.2 So dium dodecyl sulfate-polyacrylamide gel electrophoresis of glycoproteins of HL-60/AR revealed two prominent glycoproteins with molecular weights of 160,000 ± 10,000 and 110,000 ± 10,000 which were not detected in the sensitive cells. Cellular uptake and retention of daunorubicin was studied in the resistant and sensitive cells utilizing digitized video fluorescence micros copy. The sensitive cells accumulated more drug and showed at least 2-fold greater levels of brightness than the resistant cells. Studies of total intracellular accumulation, utilizing 10~6 M [14C]daunorubicin as a marker, showed a 1-h accumulation of 98 ± 20 pmol/106 cells in HL-60/AR versus 255 ±25 pmol/106 cells in HL-60. Exposure to nontoxic concentrations of the calcium channel blocker Verapamil (10~5 M)led to enhanced accumulation (175 ±8 pmol/106 cells) and retention of the drug in HL-60/AR, resulting in increased cytotoxicity in HL-60/AR. These anthra cycline-resistant leukemic cells may serve as a valuable experi mental model in studying the phenomenon of multiple drug resistance as well as strategies to circumvent it in human myeloid leukemia.
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تاریخ انتشار 2006