Expression of β1,3-N-acetylglucosaminyltransferases during differentiation of human acute myeloid leukemia cells
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The expressions of β1,3-N-acetylglucosamonyltransferase-2 and -8 (β3GnT-2, β3GnT-8),—the two main glycosyltransferases responsible for the synthesis of poly-N-acetyllactosamine (polyLacNAc) in glycans, and β3GnT-5 participating in the syntheses of sphingoglycolipids were studied in leukemia cell lines during differentiation using RT-PCR method. β3GnT-2 and β3GnT-8 distribute widely in six myeloid and monocytoid leukemia cell lines with different abundances, while β3GnT-4 was only present in NB4 cells. ATRA (all-trans retinoic acid) and dimethylsulfoxide (DMSO), which induce the differentiation of HL-60 and NB4 (two human acute myeloid leukemia cell lines) to myelocytic lineage, up-regulated these two enzymes with various degrees at 2 and 72 h of treatment. In HL-60 cells treated with ATRA, the increase of β3GnT-8 was more than β3GnT-2, while in NB4 cells treated with DMSO, the increase of β3GnT-2 was more than β3GnT-8. However, when HL-60 and NB4 were differentiated to monocytic lineage induced by phorbol 12-myristate 13-acetate the expressions of β3GnT-2 and β3GnT-8 showed no alterations or the increase of expressions was far less than those in myelocytic differentiation. By means of FITC-labeled tomato lectin affinity staining and flow-cytometry, it was found that the product of β3GnT-2 and -8, polyLacNAc was also increased on the cell surface of HL-60 and NB4 treated with ATRA or DMSO, but unchanged when treated with PMA. These results were in accordance with the up-regulation of the mRNAs of β3GnT-2 and -8. The expression of β3GnT-5, however, was not changed both in myelocytic and monocytic differentiations. The difference in the up-regulation of β3GnT-2 and -8, especially their products may become a useful index to discriminate the myelocytic and monocytic differentiation of leukemia cells.
KeywordsLeukemia cell lines Differentiation-inducers β1, 3-N-Acetylglucosaminyltransferase RT-PCR Flow-cytometric analysis
All-trans retinoic acid
Phorbol 12-myristate 13-acetate
- GlcNAc (Gn)
Reverse transcriptase-polymerase chain reaction
Fetal calf serum
N-(2-Hydroxyethyl) piperazine-N-(2-ethanesulfonic acid).
This research was supported by the grant from National Science Foundition of China (No. 30670642).
- 3.Li Z, Zhang Y, Zhao JH, Chen HL (2000) Effects of all-trans retinoic acid and phorbol ester on the surface N-linked sugar chains of HL-60 cells. China Oncol 10:19–22 (in Chinese)Google Scholar
- 7.Dong SC, Yang XP, Chen HL (1995) Effect of dibutyryl cAMP on the type and antennary number of N-glycans on the structure of human hepatocarcinoma cell line SMMC-7721. Chin J Biochem Biophys 2:103–109 (in Chinese)Google Scholar
- 8.Mizoguchi A, Takasaki S, Maeda S, Kobata A (1984) Changes in asparagines-linked sugar chains of human promyelocytic leukemia cells (HL-60) during monocytoid differentiation and myeloid differentiation. Appearance of high-mannose-type oligosacchrides in neutral fraction. J Biol Chem 259:11943–11948PubMedGoogle Scholar
- 9.Mizoguchi A, Takasaki S, Maeda S, Kobata A (1984) Changes in asparagines-linked sugar chains of human promyelocytic leukemia cells (HL-60) during monocytoid differentiation and myeloid differentiation. Decrease of high-molecular-weight oligosaccharides in acidic fraction. J Biol Chem 259:11949–11957PubMedGoogle Scholar
- 13.Ishida H, Togayachi A, Sakai T, Iwai T, Hiruma T, Sato T, Okubo R, Inaba N, Kudo T, Gotoh M, Shoda J, Tanaka N, Narimatsu H (2005) A novel β1, 3-N-acetylglucosaminyltransferase (b3Gn-T8), which synthesizes poly-N-acetyl-lactamine, is dramatically upregulated in colon cancer. FEBS Lett 579:71–78PubMedCrossRefGoogle Scholar
- 15.Fukuda M (1994) Cell surface carbohydrate: cell type-specific expression. In: Fukuda M, Hindsgaul O (eds) Molecular glycobiology. Oxford IRL Press, OxfordGoogle Scholar