Journal of Gastroenterology

, Volume 49, Issue 4, pp 702–714 | Cite as

Expression of N-acetylglucosaminyltransferase V in the subserosal layer correlates with postsurgical survival of pathological tumor stage 2 carcinoma of the gallbladder

  • Kenichiro Onuki
  • Hiroaki Sugiyama
  • Kazunori Ishige
  • Toru Kawamoto
  • Takehiro Ota
  • Shunichi Ariizumi
  • Masayuki Yamato
  • Shinichi Kadota
  • Kaoru Takeuchi
  • Akiko Ishikawa
  • Masafumi Onodera
  • Kojiro Onizawa
  • Masakazu Yamamoto
  • Eiji Miyoshi
  • Junichi Shoda
Original Article—Liver, Pancreas, and Biliary Tract



N-Acetylglucosaminyltransferase V (GnT-V), an enzyme that catalyzes the β1-6 branching of N-acetylglucosamine on asparagine-linked oligosaccharides of cellular proteins, enhances the malignant behaviors of carcinoma cells in experimental models. The aim of this study was to determine clinical significance of GnT-V expression in human pT2 gallbladder carcinoma with simple in vitro experiments.


Ninety patients with pT2 gallbladder carcinoma were included for this study. The in vitro and in vivo biological effects of GnT-V were investigated using gallbladder carcinoma cells with variable GnT-V expression levels induced by a small interfering RNA.


Of the 90 cases, 57 showed positive staining and the remaining 33 demonstrated negative staining, the subcellular localization in the 57 cases was classified into the granular-type in 31 cases and the diffuse-type in 26 cases. In 76 cases with curative resection, postsurgical survival was significantly poorer in those showing positive staining than in those showing negative staining (P = 0.028). In all of the 76 cases, postsurgical recurrence was significantly more frequent in those showing diffuse-type localization than in those showing negative staining. Experimental analyses demonstrated that the down-regulation of GnT-V expression in gallbladder carcinoma cells induced suppression of cell growth in vitro. The expression levels of GnT-V in the cells were highly correlated with the rapid in vivo growth coupled with the enhanced angiogenesis, and the tendency to form liver metastasis.


GnT-V expression in the subserosal layer of pT2 gallbladder carcinoma is correlated with the aggressiveness of the disease.


Gallbladder carcinoma N-Acetylglucosaminyltransferase V Malignant behavior Postsurgical prognosis Tumor cell biology 



Monoclonal antibody






Pathological tumor stage



Grant support: This work was supported in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology, Japan (Nos. 22390379, 22591530, 23390318, 23659659, 24390323).

Conflict of interest

The authors declare that they have no conflict of interest.


  1. 1.
    Ruckert JC, Ruckert RI, Gellert K, Hecker K, Muller JM. Surgery for carcinoma of the gallbladder. Hepatogastroenterology. 1996;43:527–33.PubMedGoogle Scholar
  2. 2.
    Cubertafond P, Gainant A, Cucchiaro G. Surgical treatment of 724 carcinomas of the gallbladder. Results of the French Surgical Association Survey. Ann Surg. 1994;219:275–80.PubMedCentralPubMedCrossRefGoogle Scholar
  3. 3.
    Oertli D, Herzog U, Tondelli P. Primary carcinoma of the gallbladder: operative experience during a 16-year period. Eur J Surg. 1993;159:415–20.PubMedGoogle Scholar
  4. 4.
    Gall FP, Kockerling F, Scheele J, Schneider C, Hohenberger W. Radical operations for carcinoma of the gallbladder: present status in Germany. World J Surg. 1991;15:328–36.PubMedCrossRefGoogle Scholar
  5. 5.
    Donohue JH, Nagorney DM, Grant CS, Tsushima K, Ilstrup DM, Adson MA. Carcinoma of the gallbladder. Does radical resection improve outcome? Arch Surg. 1990;125:237–41.PubMedCrossRefGoogle Scholar
  6. 6.
    Ouchi K, Owada Y, Matsuno S, Sato T. Prognostic factors in the surgical treatment of gallbladder carcinoma. Surgery. 1987;101:731–7.PubMedGoogle Scholar
  7. 7.
    Yamaguchi K, Chijiiwa K, Saiki S, Nishihara K, Takashima M, Kawakami K, et al. Retrospective analysis of 70 operations for gallbladder carcinoma. Br J Surg. 1997;84:200–4.PubMedCrossRefGoogle Scholar
  8. 8.
    Todoroki T, Kawamoto T, Takahashi Y, Takada Y, Koike N, Otsuka M, et al. Treatment of gallbladder cancer by radical resection. Br J Surg. 1999;86:622–7.PubMedCrossRefGoogle Scholar
  9. 9.
    Kawamoto T, Shoda J, Irimura T, Miyahara N, Furukawa M, Ueda T, et al. Expression of MUC1 mucins in the subserosal layer correlates with post-surgical prognosis of pT2 carcinoma of the gallbladder. Clin Cancer Res. 2001;7:1333–42.PubMedGoogle Scholar
  10. 10.
    Hart GW, Copeland RJ. Glycomics hits the big time. Cell. 2010;143:672–6.PubMedCentralPubMedCrossRefGoogle Scholar
  11. 11.
    Dennis JW, Laferte S, Waghorne C, Breitman ML, Kerbel RS. Beta 1–6 branching of Asn-linked oligosaccharides is directly associated with metastasis. Science. 1987;236:582–5.PubMedCrossRefGoogle Scholar
  12. 12.
    Miyoshi E, Nishikawa A, Ihara Y, Gu J, Sugiyama T, Hayashi N, et al. N-Acetylglucosaminyltransferase III and V messenger RNA levels in LEC rats during hepatocarcinogenesis. Cancer Res. 1993;53:3899–902.PubMedGoogle Scholar
  13. 13.
    Lau KS, Dennis JW. N-glycans in cancer progression. Glycobiology. 2008;18:750–60.PubMedCrossRefGoogle Scholar
  14. 14.
    Granovsky M, Fata J, Pawling J, Muller WJ, Khokaha R, Dennis JW. Supression of tumor growth and metastasis in Mgat5-deficient mice. Nat Med. 2000;6:306–12.PubMedCrossRefGoogle Scholar
  15. 15.
    Dennis JW, Nabi IR, Demetriou M. Metabolism, cell surface organization, and disease. Cell. 2009;139:1229–41.PubMedCentralPubMedCrossRefGoogle Scholar
  16. 16.
    Ihara S, Miyoshi E, Ko JH, Murata K, Nakahara S, Honke K, et al. Prometastatic effect of N-Acetylglucosaminyltransferase V is due to modification and stabilization of active matriptase by adding beta 1–6 GlcNAc branching. J Biol Chem. 2002;277:16960–7.PubMedCrossRefGoogle Scholar
  17. 17.
    Saito T, Miyoshi E, Sasai K, Nakano N, Eguchi H, Honke K, et al. A secreted type of beta 1,6-N-Acetylglucosaminyltransferase V (GnT-V) induces tumor angiogenesis without mediation of glycosylation: a novel function of GnT-V distinct from the original glycosyltransferase activity. J Biol Chem. 2002;277:17002–8.PubMedCrossRefGoogle Scholar
  18. 18.
    Murata K, Miyoshi E, Kameyama M, Ishikawa O, Kabuto T, Sasaki Y, et al. Expression of N-Acetylglucosaminyltransferase V in colorectal cancer correlates with metastasis and poor prognosis. Clin Cancer Res. 2000;6:1772–7.PubMedGoogle Scholar
  19. 19.
    Perng GS, Shoreibah M, Margitich I, Pierce M, Fregien N. Expression of N-Acetylglucosaminyltransferase V mRNA in mammalian tissues and cell lines. Glycobiology. 1994;4:867–71.PubMedCrossRefGoogle Scholar
  20. 20.
    Dosaka-Akita H, Miyoshi E, Suzuki O, Itoh T, Katoh H, Taniguchi N. Expression of N-Acetylglucosaminyltransferase V is associated with prognosis and histology in non-small cell lung cancers. Clin Cancer Res. 2004;10:1773–9.PubMedCrossRefGoogle Scholar
  21. 21.
    Ishimura H, Takahashi T, Nakagawa H, Nishimura S, Arai Y, Horikawa Y, et al. N-Acetylglucosaminyltransferase V and beta 1–6 branching N-linked oligosaccharides are associated with good prognosis of patients with bladder cancer. Clin Cancer Res. 2006;12:2506–11.PubMedCrossRefGoogle Scholar
  22. 22.
    Handerson T, Pawelek JM. Beta 1,6-branched oligosaccharides and coarse vesicles: a common, pervasive phenotype in melanoma and other human cancers. Cancer Res. 2003;63:5363–9.PubMedGoogle Scholar
  23. 23.
    Fleming ID, Cooper JS, Henson DE, Hutter RVP, Kennedy BJ, Murphy GP, O’sullivan B, Sobin LH, Yarbro JW, editors. AJCC cancer staging manual. 5th ed. Philadelphia: Lippincott-Raven; 1997.Google Scholar
  24. 24.
    Knuth A, Gabbert H, Dippold W, Klein O, Sachsse W, Bitter-Suermann D, et al. Biliary adenocarcinoma. Characterisation of three new human tumor cell lines. J Hepatol. 1985;1:579–96.PubMedCrossRefGoogle Scholar
  25. 25.
    Koike N, Todoroki T, Kawamoto T, Yoshida S, Kashiwagi H, Fukao K, et al. The invasion potentials of human biliary tract carcinoma cell lines: correlation between invasiveness and morphologic characteristics. Int J Oncol. 1998;13:1269–74.PubMedGoogle Scholar
  26. 26.
    Miyahara N, Shoda J, Kawamoto T, Furukawa M, Ueda T, Todoroki T, et al. Expression of UDP-N-Acetyl-alpha-d-galactosamine-polypeptide N-Acetylgalactosaminyltransferase isozyme 3 in the subserosal layer correlates with postsurgical survival of pathological tumor stage 2 carcinoma of the gallbladder. Clin Cancer Res. 2004;10:2090–9.PubMedCrossRefGoogle Scholar
  27. 27.
    Weidner N, Semple JP, Welch WR, Folkman J. Tumor angiogenesis and metastasis-correlation in invasive breast carcinoma. N Engl J Med. 1991;324:1–8.PubMedCrossRefGoogle Scholar
  28. 28.
    Inamori K, Gu J, Ohira M, Kawasaki A, Nakamura Y, Nakagawa T, et al. High expression of N-Acetylglucosaminyltransferase V in favorable neuroblastomas: Involvement of its effect on apotosis. FEBS Lett. 2006;580:627–32.PubMedCrossRefGoogle Scholar
  29. 29.
    Nabekura T, Otsu M, Nagasawa T, Nakauchi H, Onodera M. Potent vaccine therapy with dendritic cells genetically modified by the gene-silencing-resistant retroviral vector GCDNsap. Mol Ther. 2006;13:301–9.PubMedCrossRefGoogle Scholar
  30. 30.
    Suzuki A, Obi K, Urabe T, Hayakawa H, Yamada M, Kaneko S, et al. Feasibility of ex vivo gene therapy for neurological disorders using the new retroviral vector GCDNsap packaged in the vesicular stomatitis virus G protein. J Neurochem. 2002;82:953–60.PubMedCrossRefGoogle Scholar
  31. 31.
    Taniguchi N, Miyoshi E, Ko JH, Ikeda Y, Ihara Y. Implication of N-Acetylglucosaminyltransferases III and V in cancer: gene regulation and signaling mechanism. Biochim Biophys Acta. 1999;1455:287–300.PubMedCrossRefGoogle Scholar
  32. 32.
    Ichihara H, Funamoto K, Matsushita T, Matsumoto Y, Ueoka R. Histological bioanalysis for therapeutic effects of hybrid liposomes on the hepatic metastasis of colon carcinoma in vivo. Int J Pharm. 2010;394:174–8.PubMedCrossRefGoogle Scholar
  33. 33.
    Tsukada K, Hatakeyama K, Kurosaki I, Uchida K, Shirai Y, Muto T, et al. Outcome of radical surgery for carcinoma of the gallbladder according to the TNM stage. Surgery. 1996;120:816–21.PubMedCrossRefGoogle Scholar
  34. 34.
    Matsushita Y, Cleary KR, Ota DM, Hoff SD, Irimura T. Syalyl-dimetric Lewis-X antigen expressed on mucin-like glycoproteins in colorectal cancer metastasis. Lab Invest. 1990;63:780–91.PubMedGoogle Scholar
  35. 35.
    Kawamoto T, Shoda J, Miyahara N, Suzuki H, Furukawa M, Todoroki T, et al. Expression of MUC1 recognized by a monoclonal antibody MY.1E12 is a useful biomarker for tumor aggressiveness of carcinoma of the gallbladder. Clin Exp Metastasis. 2004;21:353–62.Google Scholar
  36. 36.
    Zhang K, Baeckstrom D, Brevinge H, Hansson GC. Secreted MUC1 mucins lacking their cytoplasmic part and carrying sialylLewis a and x epitopes from a tumor cell line and sera of colon carcinoma patients can inhibit HL-60 leukocyte adhesion to E-selectin-expressing endothelial cells. J Cell Biochem. 1996;60:538–49.PubMedCrossRefGoogle Scholar
  37. 37.
    Treon SP, Maimonis P, Bua D, Young G, Raje N, Mollick J, et al. Elevated soluble MUC1 levels and decreased anti-MUC1 antibody levels in patients with multiple melanoma. Blood. 2000;96:3147–53.PubMedGoogle Scholar
  38. 38.
    Thathiah A, Blobel CP, Carson DD. Tumor necrosis factor-α converting enzyme/ADAM 17 mediates MUC1 shedding. J Biol Chem. 2003;278:3386–94.PubMedCrossRefGoogle Scholar
  39. 39.
    Taatjes DJ, Roth J, Weinstein J, Paulson JC, Shaper NL, Shaper JH. Codistribution of galactosyl- and sialyltransferase: reorganization of trans Golgi apparatus elements in hepatocytes in intact liver and cell culture. Eur J Cell Biol. 1987;44:187–94.PubMedGoogle Scholar
  40. 40.
    Pierce M, Buckhaults P, Chen L, Fregien N. Regulation of N-acetylglucosaminyltransferase V and Asn-linked oligosaccharide beta (1,6) branching by a growth factor signaling pathway and effects on cell adhesion and metastatic potential. Glycoconj J. 1997;14:623–30.PubMedCrossRefGoogle Scholar
  41. 41.
    Ujita M, McAuliffe J, Hindsgaul O, Sasaki K, Fukuda MN, Fukuda M. Poly-N-acetyllactosamine synthesis in branched N-glycans is controlled by complemental branch specificity of I-extension enzyme and beta1, 4-galactosyltransferase I. J Biol Chem. 1999;274:16717–26.PubMedCrossRefGoogle Scholar
  42. 42.
    Saitoh O, Wang WC, Lotan R, Fukuda M. Differential glycosylation and cell surface expression of lysosomal membrane glycoproteins in sublines of a human colon cancer exhibiting distinct metastatic potentials. J Biol Chem. 1992;267:5700–11.PubMedGoogle Scholar
  43. 43.
    Nakamori S, Kameyama M, Imaoka S, Furukawa H, Ishikawa O, Sasaki Y, et al. Increased expression of sialyl Lewisx antigen correlates with poor survival in patients with colorectal carcinoma: clinicopathological and immunohistochemical study. Cancer Res. 1993;53:3632–7.PubMedGoogle Scholar
  44. 44.
    Shimodaira K, Nakayama J, Nakamura N, Hasebe O, Katsuyama T, Fukuda M. Carcinoma-associated expression of core 2 beta-1,6-N-Acetylglucosaminyltransferase gene in human colorectal cancer: role of O-glycans in tumor progression. Cancer Res. 1997;57:5201–6.PubMedGoogle Scholar
  45. 45.
    Nakahara S, Saito T, Kondo N, Moriwaki K, Noda K, Ihara S, et al. A secreted type of β1,6 N-acetylglucosaminyltransferase V (GnT-V), a novel angiogenesis inducer, is regulated by γ-secretase. FASEB J. 2006;20:2451–9.PubMedCrossRefGoogle Scholar
  46. 46.
    Dennis JW. Effects of swainsonine and polycytidylic acid on murine tumor cell growth and metastasis. Cancer Res. 1986;46:5131–6.PubMedGoogle Scholar
  47. 47.
    Goss PE, Reid CL, Bailey D, Dennis JW. Phase IB clinical trial of the oligosaccharide processing inhibitor swainsonone in patients with advanced malignancies. Clin Cancer Res. 1997;3:1077–86.PubMedGoogle Scholar

Copyright information

© Springer Japan 2013

Authors and Affiliations

  • Kenichiro Onuki
    • 1
  • Hiroaki Sugiyama
    • 2
  • Kazunori Ishige
    • 2
  • Toru Kawamoto
    • 1
  • Takehiro Ota
    • 1
  • Shunichi Ariizumi
    • 1
  • Masayuki Yamato
    • 3
  • Shinichi Kadota
    • 4
  • Kaoru Takeuchi
    • 4
  • Akiko Ishikawa
    • 5
  • Masafumi Onodera
    • 6
  • Kojiro Onizawa
    • 7
  • Masakazu Yamamoto
    • 1
  • Eiji Miyoshi
    • 5
  • Junichi Shoda
    • 8
  1. 1.Department of Surgery, Institute of GastroenterologyTokyo Women’s Medical UniversityTokyoJapan
  2. 2.Department of Gastroenterology, Division of Biomedical Science, Faculty of MedicineUniversity of TsukubaTsukubaJapan
  3. 3.Institute of Advanced Biomedical Engineering and ScienceTokyo Women’s Medical UniversityTokyoJapan
  4. 4.Department of Infection Biology, Division of Biomedical Science, Faculty of MedicineUniversity of TsukubaTsukubaJapan
  5. 5.Division of Health SciencesOsaka University Graduate School of MedicineSuitaJapan
  6. 6.Department of GeneticsNational Research Institute for Child Health and DevelopmentTokyoJapan
  7. 7.Department of Oral and Maxillofacial Surgery, Division of Biomedical Science, Faculty of MedicineUniversity of TsukubaTsukubaJapan
  8. 8.Medical Science, Division of Clinical Science, Faculty of MedicineUniversity of TsukubaTsukubaJapan

Personalised recommendations