Alterations in glycosyltransferases during myeloid and monocytoid differentiation of HL-60 cells.
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 GM(3). The activity alterations of N -acetylglucosaminyltransferase III and V (GnT-III, GnT-V) as well as alpha-1,6-fucosyl-tranferase (alpha1,6 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 alpha1,6 Fuc T assay. It was observed that after 3 days, all three enzymes decreased in HL-60 cells induced by 1 micromol/l ATRA and 0.6 mmol/l n-BA, while GnT-III and alpha1,6 Fuc T increased, but GnT-V still decreased after induction by 1% DMSO. GnT-V and alpha1,6 Fuc T declined, while GnT-III was elevated after induction by 0.1 micromol/l PMA for 3 days. In contrast, GnT-III increased after the treatment with 50 micromol/l GM(3)for 3 or 6 days, but GnT-V was not appreciably changed and alpha1,6 FucT was elevated after 6 days of GM(3)treatment. 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.
Liu
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AH
Key Laboratory of Glycoconjugate Research, Ministry of Public Health, Department of Biochemistry, Shanghai Medical University, Shanghai, 200032, P.R. China.
Liu
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Li
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Gu
J X
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Chen
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HL
D000970:Antineoplastic Agents / Q000494:pharmacology; D002454:Cell Differentiation; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004790:Enzyme Induction; D005647:Fucosyltransferases / Q000378:metabolism; D018922:HL-60 Cells / Q000187:drug effects / Q000201:enzymology*; D006801:Humans; D009000:Monocytes / Q000166:cytology / Q000187:drug effects; D017351:N-Acetylglucosaminyltransferases / Q000187:drug effects / Q000378:metabolism*; D014212:Tretinoin / Q000494:pharmacology
D000970:Antineoplastic Agents; D014212:Tretinoin; D005647:Fucosyltransferases; D017351:N-Acetylglucosaminyltransferases; C054599:beta-1,4-mannosyl-glycoprotein beta-1,4-N-acetylglucosaminyltransferase; C052254:alpha-1,6-mannosylglycoprotein beta 1,6-N-acetylglucosaminyltransferase; D004121:Dimethyl Sulfoxide