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Expression of β1,3-N-acetylglucosaminyltransferases during differentiation of human acute myeloid leukemia cells

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Abstract

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.

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Abbreviations

ATRA:

All-trans retinoic acid

DMSO:

Dimethylsulfoxide

PMA:

Phorbol 12-myristate 13-acetate

PolyLacNAc:

Poly-N-acetyllactosamine

Gal:

Galactose

GlcNAc (Gn):

N-Acetylglucosamine

GnT:

N-Acetylglucosaminyltransferase

Man:

Mannose

RT-PCR:

Reverse transcriptase-polymerase chain reaction

FITC:

Fluorescein isothiocynate

FCS:

Fetal calf serum

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

HEPES:

N-(2-Hydroxyethyl) piperazine-N-(2-ethanesulfonic acid).

PBS:

Phosphate-buffered saline

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Acknowledgment

This research was supported by the grant from National Science Foundition of China (No. 30670642).

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Medical School of Soochow University, Suzhou, 215123, China

    Hao Qiu, Shi-Liang Wu, Xiang-Hong Guo & Huan-Ping Zhang

  2. Jiangsu Institute of Hematology, The First Affiliated Hospital of Soochow University, Suzhou, 215006, China

    Hong-Jie Shen

  3. Key Laboratory of Glycoconjugate Research, Department of Biochemistry and Molecular Biology, Shanghai Medical College, Fudan University, Shanghai, 200032, China

    Hui Li Chen

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  1. Hao Qiu
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  2. Shi-Liang Wu
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Correspondence to Hui Li Chen.

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Hao Qiu and Shi-Liang Wu contributed equally to this article.

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Qiu, H., Wu, SL., Guo, XH. et al. Expression of β1,3-N-acetylglucosaminyltransferases during differentiation of human acute myeloid leukemia cells. Mol Cell Biochem 358, 131–139 (2011). https://doi.org/10.1007/s11010-011-0928-9

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