Glycoconjugate Journal

, Volume 25, Issue 3, pp 225–235 | Cite as

Transcriptional regulation of the fucosyltransferase VI gene in hepatocellular carcinoma cells

  • Koji HigaiEmail author
  • Noriko Miyazaki
  • Yutaro Azuma
  • Kojiro Matsumoto


The α1,3-fucosyltransferase VI (FUT VI) protein is a key enzyme for synthesis of sialyl Lewis X and Lewis X in epithelial cells. Despite its importance, how FUT VI expression is regulated has not previously been elucidated. In this work, we examined transcriptional regulation of the FUT VI gene in hepatocellular carcinoma HepG2 cells. 5′-Rapid amplification of cDNA ends analysis revealed transcription start sites of FUT VI in HepG2 cells at +65 and +278 nucleotides (nt) downstream of the position registered in the Data Base of Human Transcription Start Sites. We determined promoter regions for FUT VI in HepG2 cells using a luciferase reporter gene assay. The promoter activities of constructs located 5′-upstream of the transcription start site decreased when the −186 to −156 and −56 to −19 nt regions were deleted. Site-directed mutagenesis of these regions revealed that two hepatocyte nuclear factor-4α (HNF-4α) and one octamer binding transcription factor-1 (Oct-1) binding sites are essential for FUT VI transcription. Furthermore, transient over-expression of HNF-4α but not Oct-1 enhanced both FUT VI promoter activities and FUT VI mRNA levels in HuH-7 cells. These results suggest that two defined regions in the 5′-flanking region of the FUT VI transcription start site are critical for FUT VI transcription in HepG2 cells.


Fucosyltransferase VI Hepatocyte nuclear factor HepG2 cells Transcriptional regulation 



This research was partially supported by the Ministry of Education, Science, Sports and Culture, Grant-in-Aid for Young Scientists (B), 19790398, 2007 and by the “Open Research Center” Project.


  1. 1.
    Lowe, J.B.: Glycan-dependent leukocyte adhesion and recruitment in inflammation. Curr. Opin. Cell Biol. 15, 531–538 (2003)PubMedCrossRefGoogle Scholar
  2. 2.
    Kannagi, R., Izawa, M., Koike, T., Miyazaki, K., Kimura, N.: Carbohydrate-mediated cell adhesion in cancer metastasis and angiogenesis. Cancer Sci. 95, 377–384 (2004)PubMedCrossRefGoogle Scholar
  3. 3.
    Kannagi, R.: Molecular mechanism for cancer-associated induction of sialyl Lewis X and sialyl Lewis A expression. The Warburg effect revisited. Glycoconj J 20, 353–364 (2004)PubMedCrossRefGoogle Scholar
  4. 4.
    Kukowska-Latallo, J.F., Larsen, R.D., Nair, R.P., Lowe, J.B.: A cloned human cDNA determines expression of a mouse stage-specific embryonic antigen and the Lewis blood group a1,3/1,4-fucosyltransferase. Genes Dev 4, 1288–303 (1990)PubMedCrossRefGoogle Scholar
  5. 5.
    Lowe, J.B., Kukowska-Latallo, J.F., Nair, R.P., Larsen, R.D., Marks, R.M., Macher, B.A., Kelly, R.J., Ernst, L.K.: Molecular cloning of a human fucosyltransferase gene that determines expression of the Lewis X and VIM-2 epitopes but not ELAM-1-dependent cell adhesion. J Biol Chem 266, 17467–17477 (1991)PubMedGoogle Scholar
  6. 6.
    Weston, B.W., Nair, R.P., Larsen, R.D., Lowe, J.B.: Isolation of a novel human a(1,3)fucosyltransferase gene and molecular comparison to the human Lewis blood group a1,3/1,4-fucosyltransferase gene. Syntenic, homologous, nonallelic genes encoding enzymes with distinct acceptor substrate specificities. J. Biol. Chem. 267, 4152–4160 (1992)PubMedGoogle Scholar
  7. 7.
    Weston, B.W., Smith, P.L., Kelly, R.J., Lowe, J.B.: Molecular cloning of a fourth member of a human a1,3-fucosyltransferase gene family. Multiple homologous sequences that determine expression of the Lewis X, sialyl Lewis X, and difucosyl sialyl Lewis X epitopes. J. Biol. Chem. 267, 24575–24584 (1992)PubMedGoogle Scholar
  8. 8.
    Koszdin, K.L., Bowen, B.R.: The cloning and expression of a human a1,3 fucosyltransferase capable of forming the E-selectin ligand. Biochem. Biophys. Res. Commun. 187, 152–157 (1992)PubMedCrossRefGoogle Scholar
  9. 9.
    Natsuka, S., Gersten, K.M., Zenita, K., Kannagi, R., Lowe, J.B.: Molecular cloning of a cDNA encoding a novel human leukocyte a1,3-fucosyltransferase capable of synthesizing the sialyl Lewis X determinant. J Biol Chem 269, 16789–16794 (1994)PubMedGoogle Scholar
  10. 10.
    Sasaki, K., Kurata, K., Funayama, K., Nagata, M., Watanabe, E., Ohta, S., Hanai, N., Nishi, T.: Expression cloning of a novel a1,3-fucosyltransferase that is involved in biosynthesis of the sialyl Lewis X carbohydrate determinants in leukocytes. J. Biol. Chem. 269, 14730–14737 (1994)PubMedGoogle Scholar
  11. 11.
    Kaneko, M., Kudo, T., Iwasaki, H., Ikehara, Y., Nishihara, S., Nakagawa, S., Sasaki, K., Shiina, T., Inoko, H., Saitou, N., Narimatsu, H.: a1,3-Fucosyltransferase IX (Fuc-TIX) is very highly conserved between human and mouse. Molecular cloning, characterization and tissue distribution of human Fuc-TIX. FEBS Lett 452, 237–242 (1999)PubMedCrossRefGoogle Scholar
  12. 12.
    Shinoda, K., Tanahashi, E., Fukunaga, K., Ishida, H., Kiso, M.: Detailed acceptor specificities of human a1,3-fucosyltransferases, Fuc-TVII and Fuc-TVI. Glycoconj. J. 15, 969–974 (1998)PubMedCrossRefGoogle Scholar
  13. 13.
    Huang, M.C., Laskowska, A., Vestweber, D., Wild, M.K.: The a1,3-fucosyltransferase Fuc-TIV, but not Fuc-TVII, generates sialyl Lewis X-like epitopes preferentially on glycolipids. J. Biol. Chem. 277, 4778–4795 (2002)CrossRefGoogle Scholar
  14. 14.
    Kimura, H., Shinya, N., Nishihara, S., Kaneko, M., Irimura, T., Narimatsu, H.: Distinct substrate specificities of five human a1,3-fucosyltransferases for in vivo synthesis of the sialyl Lewis X and Lewis X epitopes. Biochem. Biophys. Res. Commun. 237, 131–137 (1997)PubMedCrossRefGoogle Scholar
  15. 15.
    Borsig, L., Imbach, T., Hochli, M., Berger, E.G.: a1,3-Fucosyltransferase VI is expressed in HepG2 cells and codistributed with a1,4-galactosyltransferase I in the Golgi apparatus and monensin-induced swollen vesicles. Glycobiology 9, 1273–1280 (1999)PubMedCrossRefGoogle Scholar
  16. 16.
    Higai, K., Shibukawa, K., Muto, S., Matsumoto, K.: Targeted proteo-glycomics analysis of sialyl Lewis X antigen expressing glycoproteins secreted by human hepatoma cell line. Anal. Sci. 19, 85–92 (2003)PubMedCrossRefGoogle Scholar
  17. 17.
    Wang, Q.Y., Wu, S.L., Chen, J.H., Liu, F., Chen, H.L.: Expressions of Lewis antigens in human non-small cell pulmonary cancer and primary liver cancer with different pathological conditions. J. Exp. Clin. Cancer Res. 22, 431–440 (2003)PubMedGoogle Scholar
  18. 18.
    Yazawa, S., Madiyalakan, R., Izawa, H., Asao, T., Furukawa, K., Matta, K.L.: Cancer-associated elevation of a1,3-l-fucosyltransferase activity in human serum. Cancer 62, 516–520 (1988)PubMedCrossRefGoogle Scholar
  19. 19.
    Kuzmits, R., Aiginger, P.: Evaluation of serum fucosyltransferases in malignancy. Eur. J. Cancer Clin. Oncol. 20, 667–672 (1984)PubMedCrossRefGoogle Scholar
  20. 20.
    Madiyalakan, R., Yazawa, S., Barlow, J.J., Matta, K.L.: Elevated serum a1,3-l-fucosyltransferase activity with synthetic low molecular weight acceptor in human ovarian cancer. Cancer Lett. 30, 201–205 (1986)PubMedCrossRefGoogle Scholar
  21. 21.
    Tanaka, S., Yazawa, S., Noguchi, K., Nishimura, T., Miyanaga, K., Kochibe, N., Poland, D.C., van Dijk, W., Matta, K.L.: Molecular analysis of plasma a1,3-fucosyltransferase deficiency and development of the methods for its genotyping. Exp. Clin. Immunogenet. 18, 1–12 (2001)PubMedCrossRefGoogle Scholar
  22. 22.
    Fraser, J.D., Martinez, V., Straney, R., Briggs, M.R.: DNA binding and transcription activation specificity of hepatocyte nuclear factor 4. Nucleic Acids Res. 26, 2702–2707 (1998)PubMedCrossRefGoogle Scholar
  23. 23.
    Groenen, M.A., Dijkhof, R.J., van der Poel, J.J., van Diggelen, R., Verstege, E.: Multiple octamer binding sites in the promoter region of the bovine as2-casein gene. Nucleic Acids Res. 20, 4311–4318 (1992)PubMedCrossRefGoogle Scholar
  24. 24.
    Kanoh, A., Ota, M., Narimatsu, H., Irimura, T.: Expression levels of FUT6 gene transfected into human colon carcinoma cells switch two sialyl-Lewis X-related carbohydrate antigens with distinct properties in cell adhesion. Biochem. Biophys. Res. Commun. 303, 869–901 (2003)CrossRefGoogle Scholar
  25. 25.
    Dabrowska, A., Baczynska, D., Widerak, K., Laskowska, A., Ugorski, M.: Promoter analysis of the human a1,3/4-fucosyltransferase gene (FUT III). Biochim. Biophys. Acta 1731, 66–73 (2005)PubMedGoogle Scholar
  26. 26.
    Taniguchi, A., Suga, R., Matsumoto, K.: Expression and transcriptional regulation of the human a1,3-fucosyltransferase 4 (FUT4) gene in myeloid and colon adenocarcinoma cell lines. Biochem. Biophys. Res. Commun. 273, 370–376 (2000)PubMedCrossRefGoogle Scholar
  27. 27.
    Hiraiwa, N., Yabuta, T., Yoritomi, K., Hiraiwa, M., Tanaka, Y., Suzuki, T., Yoshida, M., Kannagi, R.: Transactivation of the fucosyltransferase VII gene by human T-cell leukemia virus type 1 Tax through a variant cAMP-responsive element. Blood 101, 3615–3621 (2003)PubMedCrossRefGoogle Scholar
  28. 28.
    Verrijzer, C.P., Van der Vliet, P.C.: POU domain transcription factors. Biochim. Biophys. Acta 1173, 1–21 (1993)PubMedGoogle Scholar
  29. 29.
    Ryan, A.K., Rosenfeld, M.G.: POU domain family values: flexibility, partnerships, and developmental codes. Genes Dev. 11, 1207–1225 (1997)PubMedCrossRefGoogle Scholar
  30. 30.
    Sytina, E.V., Pankratova, E.V.: Transcription factor Oct-1: plasticity and multifunctionality. Mol. Biol. (Moscow) 37, 637–648 (2003)CrossRefGoogle Scholar
  31. 31.
    Schrem, H., Klempnauer, J., Borlak, J.: Liver-enriched transcription factors in liver function and development. Part I: the hepatocyte nuclear factor network and liver-specific gene expression. Pharmacol. Rev. 54, 129–158 (2002)PubMedCrossRefGoogle Scholar
  32. 32.
    Ktistaki, E., Talianidis, I.: Chicken ovalbumin upstream promoter transcription factors act as auxiliary cofactors for hepatocyte nuclear factor 4 and enhance hepatic gene expression. Mol. Cell Biol. 17, 2790–2797 (1997)PubMedGoogle Scholar
  33. 33.
    Kieback, D.G., Runnebaum, I.B., Moebus, V.J., Kreienberg, R., McCamant, S.K., Edwards, C.L., Jones, L.A., Tsai, M.J., O'Malley, B.W.: Chicken ovalbumin upstream promoter transcription factor (COUP-TF): an orphan steroid receptor with a specific pattern of differential expression in human ovarian cancer cell lines. Gynecol. Oncol. 51, 167–170 (1993)PubMedCrossRefGoogle Scholar
  34. 34.
    Wu, Q., Li, Y., Liu, R., Agadir, A., Lee, M.O., Liu, Y., Zhang, X.: Modulation of retinoic acid sensitivity in lung cancer cells through dynamic balance of orphan receptors nur77 and COUP-TF and their heterodimerization. EMBO J 16, 1656–1669 (1997)PubMedCrossRefGoogle Scholar
  35. 35.
    Navab, R., Gonzalez-Santos, J.M., Johnston, M.R., Liu, J., Brodt, P., Tsao, M.S., Hu, J.: Expression of chicken ovalbumin upstream promoter–transcription factor II enhances invasiveness of human lung carcinoma cells. Cancer Res. 64, 5097–5105 (2004)PubMedCrossRefGoogle Scholar
  36. 36.
    Kieback, D.G., Levi, T., Kohlberger, P., Fiedrich, U., Press, M.F., Rosenthal, H.E., Mobus, V.J., Runnebaum, I.B., Tong, X.W., Tsai, M.J.: Chicken ovalbumin upstream promoter–transcription factor (COUP-TF) expression in human endometrial cancer cell lines. Anticancer Res 16, 3371–3376 (1996)PubMedGoogle Scholar
  37. 37.
    Shibata, H., Ando, T., Suzuki, T., Kurihara, I., Hayashi, K., Hayashi, M., Saito, I., Murai, M., Saruta, T.: COUP-TFI expression in human adrenocortical adenomas: possible role in steroidogenesis. J. Clin. Endocrinol. Metab. 83, 4520–4523 (1998)PubMedCrossRefGoogle Scholar
  38. 38.
    Wang, Q., Bai, Z.L., Xuan, L., Hou, L., Zhang, B.: Inhibitory role of transcription factor COUP-TFII in expression of hTERT in HeLa cells. Chin. Med. Sci. J. 19, 157–163 (2004)PubMedGoogle Scholar
  39. 39.
    Wang, Q., Bai, Z., Li, X., Hou, L., Zhang, B.: The evidences of human orphan receptor COUP-TFII inhibiting telomerase activity through decreasing hTERT transcription. Cancer Lett. 214, 81–90 (2004)PubMedCrossRefGoogle Scholar
  40. 40.
    Azuma, Y., Murata, M., Matsumoto, K.: Alteration of sugar chains on α1-acid glycoprotein secreted following cytokine stimulation of HuH-7 cells in vitro. Clin. Chim. Acta 294, 93–103 (2000)PubMedCrossRefGoogle Scholar
  41. 41.
    Higai, K., Miyazaki, N., Azuma, Y., Matsumoto, K.: Interleukin-1b induces sialyl Lewis X on hepatocellular carcinoma HuH-7 cells via enhanced expression of ST3Gal IV and FUT VI gene. FEBS Lett. 580, 6069–6075 (2006)PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • Koji Higai
    • 1
    Email author
  • Noriko Miyazaki
    • 1
  • Yutaro Azuma
    • 1
  • Kojiro Matsumoto
    • 1
  1. 1.Department of Clinical Chemistry, School of Pharmaceutical SciencesToho UniversityChibaJapan

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