<b>ISOLATION AND CHARACTERIZATION OF CLONES OF A HUMAN LEUKEMIA CELL LINE (HL-60 CELLS) RESISTANT TO 1α,25-DIHYDROXYVITAMIN D<sub>3</sub> AND DIMETHYL </b><b>SULFOXIDE </b>

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  • SENOO HARUKI
    Department of Anatomy, Faculty of Medicine, Tokyo Medical and Dental University
  • WAKE KENJIRO
    Department of Anatomy, Faculty of Medicine, Tokyo Medical and Dental University
  • MOMOI TAKASHI
    Division of Pathological Chemistry, National Center for Nervous, Mental and Muscular Disorders
  • YOSHIKURA HIROSHI
    Department of Bacteriology, Faculty of Medicine, University of Tokyo

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We have previously demonstrated that HL-60 cells pretreated with 1α,25-dihydroxyvitamin D3 at the stage of commitment were induced to differentiate into macrophage- like cells by dimethyl sulfoxide (OMSO) at the stage of promotion (15). To investigate the effect of OMSO on the differentiation of HL-60 cells induced by 1α,25-dihydroxyvitamin D3, three variant clones (ORHL-1, ORHL-2 and ORHL-3 cells) resistant to 1α,25-dihydroxyvitamin D3 were iso1α,ted from a OMSO-resistant cell clone (ORHL) ofHL-60 cells. These variant clones were insusceptible to 1α,25- dihydroxyvitamin D3 or OMSO, when based on the criteria of cell growth, expression of antigens for human monocyte specific (OKMS) or human granulocyte specific (80H5) monoclona1α,ntibodies, NBT-reducing activity, phagocytosis, nonspecific esterase activity, and cathepsin Band 1α,ctivities. The variant clones, ORHL-1 and ORHL-3, which were insusceptible to 1α,25-dihydroxyvitamin D3 or OMSO alone, were induced to differentiate into macrophage-like cells on simultaneous treatment with 1α,25-dihydroxyvitamin D3 and OMSO. Furthermore, ORHL-1 but not ORHL-3 differentiated into macrophage-like cells on I-day treatment with 1α,,25-dihydroxyvitamin D3 followed by 3-day treatment with OMSO. The other variant, ORHL-2, did not differentiate with any combination of 1α,25-dihydroxyvitamin D3 and OMSO. These results suggest that OMSO promotes the differentiation of HL-60 cells initiated by 1α,,25-dihydroxyvitamin D3 into macrophages at various stages of differentiation.

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