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

  • Hao Qiu
  • Shi-Liang Wu
  • Xiang-Hong Guo
  • Hong-Jie Shen
  • Huan-Ping Zhang
  • Hui Li Chen
Article

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.

Keywords

Leukemia cell lines Differentiation-inducers β1, 3-N-Acetylglucosaminyltransferase RT-PCR Flow-cytometric analysis 

Abbreviation

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

Notes

Acknowledgment

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

References

  1. 1.
    Li Z, Liu AH, Liu F, Chen HL (1998) Modification of pentasaccharide core of surface N-glycans during differentiation of HL-60 cells. Leukemia Res 22:727–734CrossRefGoogle Scholar
  2. 2.
    Qiu H, Guo XH, Mo JH, Jin MF, Wu SL, Chen HL (2006) Expressions of polypeptide: N-acetylgalactosaminyltransferase in leukemia cell lines during 1,25-dihydroxyvitamin D3 induced differentiation. Glycoconj J 23:573–582CrossRefGoogle Scholar
  3. 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
  4. 4.
    Liu AH, Liu F, Li Z, Gu JX, Chen HL (1998) Alterations in glycosyltransferases during myeloid and monocytoid differentiation of HL-60 cells. Cell Biol Int 18:545–550CrossRefGoogle Scholar
  5. 5.
    Zhao JH, Zhang Y, Zhang XY, Chen HL (2003) Alteration in the expression of early stage processing enzymes of N-glycan during myeloid and monocytoid differentiation of HL-60 cells. Leukemia Res 27:599–605CrossRefGoogle Scholar
  6. 6.
    Chen HL, Dong SC, Ju TZ (1995) Effect of retinoic acid on the structure of N-glycans on the surface of human hepatocarcinoma cells and its enzymatic mechnism. J Cancer Res Clin Oncol 121:397–401PubMedCrossRefGoogle Scholar
  7. 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. 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. 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
  10. 10.
    Lee N, Wang WC, Fukuda M (1990) Granulocytic differentiation of HL-60 cells is associated with increase of poly-N-acetyllactosamine in Asn-linked oligo-saccharides attached to human lysosomal membrane glycoprotein. J Biol Chem 265:20476–20487PubMedGoogle Scholar
  11. 11.
    Seko A, Yamashita K (2008) Activation of β1, 3-N-Acetyl-glucosaminyltransferase-2 (β3Gn-T2) by βGn-T8: Possible involvement of 3Gn-T8 in increasing poly-N-acetyllactosamine chains in diffrentiated HL-60 cells. J Biol Chem 283:33094–33100PubMedCrossRefGoogle Scholar
  12. 12.
    Narimatsu H (2006) Human glycogene cloning: focus on β3-glycosyltransferase and β4-glycosyltransferase families. Curr Opin Struct Biol 16:567–575PubMedCrossRefGoogle Scholar
  13. 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
  14. 14.
    Liu AH, Li XB, Gu JX, Chen HL (1997) Regulatory effect of all-trans retinoic acid (ATRA) on the activities of N-acetylglucosaminyl Transferase (GnT) III, IV. Acta Biochim Biophys Sin 29:116–121 (in Chinese)PubMedGoogle Scholar
  15. 15.
    Fukuda M (1994) Cell surface carbohydrate: cell type-specific expression. In: Fukuda M, Hindsgaul O (eds) Molecular glycobiology. Oxford IRL Press, OxfordGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC. 2011

Authors and Affiliations

  • Hao Qiu
    • 1
  • Shi-Liang Wu
    • 1
  • Xiang-Hong Guo
    • 1
  • Hong-Jie Shen
    • 2
  • Huan-Ping Zhang
    • 1
  • Hui Li Chen
    • 3
  1. 1.Department of Biochemistry and Molecular BiologyMedical School of Soochow UniversitySuzhouChina
  2. 2.Jiangsu Institute of HematologyThe First Affiliated Hospital of Soochow UniversitySuzhouChina
  3. 3.Key Laboratory of Glycoconjugate Research, Department of Biochemistry and Molecular BiologyShanghai Medical College, Fudan UniversityShanghaiChina

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