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Keywords:

  • human promyelocytic leukemia cell HL-60;
  • myeloid differentiation;
  • monocytoid differentiation;
  • N -acetylglucosaminyltransferase;
  • α-1,6-fucosyltransferase

HL-60, a human promyelocytic leukemia cell line, can be differentiated to myeloid lineage by all- trans retinoic acid (ATRA), dimethylsulfoxide (DMSO) and n -butyric acid (n -BA), or to monocytoid(monocytic/macrophagic) lineage by phorbol-12-myristate-13-acetate (PMA) and ganglioside GM3. The activity alterations of N -acetylglucosaminyltransferase III and V (GnT-III, GnT-V) as well as α-1,6-fucosyl-tranferase (α1,6 Fuc T) were studied during the differentiation of HL-60 cells by the above-mentioned five inducers using the fluorescence (PA)-labeled glycan-HPLC method for GnT assays and biotin-labeled glycan-LCA affinity chromatography combined with the HRP-avidin colorimetric method for α1,6 Fuc T assay. It was observed that after 3 days, all three enzymes decreased in HL-60 cells induced by 1μmol/l ATRA and 0.6mmol/l n-BA, while GnT-III and α1,6 Fuc T increased, but GnT-V still decreased after induction by 1% DMSO. GnT-V and α1,6 Fuc T declined, while GnT-III was elevated after induction by 0.1μmol/l PMA for 3 days. In contrast, GnT-III increased after the treatment with 50μmol/l GM3for 3 or 6 days, but GnT-V was not appreciably changed and α1,6 FucT was elevated after 6 days of GM3treatment. It may be concluded that the decrease of GnT-V is the common change in myeloid differentiation and the increase of GnT-III is the general alteration in monocytoid differentiation. The changes in the activities of glycosyltransferases were consistent with the structural changes in surface N -glycans previously found in our laboratory, i.e. that the antennary number of N -glycans decreased during myeloid differentiation by ATRA, and the amount of bisecting GlcNAc in N -glycans increased during monocytoid differentiation by PMA.