Glycoantigen Biosyntheses of Human Hepatoma and Colon Cancer Cells are Dependent on Different N-Acetylglucosaminyltransferase-III and -V Activities

  • Kim, Cheorl-Ho (National Research Laboratory for Glycobiology and Department of Biochemistry and Molecular Biology, Dongguk University College of Oriental Medicine)
  • Published : 2004.10.01

Abstract

UDP-N-Acetylglucosamine(GlcNAc):$\beta$1,4-D-mannoside$\beta$-l ,4N-acetylglucosaminyltransferase-III (GnT-III) and UDP-N-GlcNAc:$\alpha$-6-D-mannosid$\beta$-1,6N-acetylglucosaminyltransferase-V(GnT - V) activities were determined in human hepatoma cell lines and metastatic colon cancer cells, and their activities were compared with those of normal liver cells and fetal hepatocytes. GnT-III activities were higher than those of GnT-V in hepatic carcinoma cells. When the two enzyme activities were assayed in highly metastatic colon cancer cells, GnT - V activities were much higher than those of GnT-III. When GlcN, GlcN-biant-PA and UDP-GlcNAc were used as substrates, the enzymes displayed different kinetic properties between hepatic and colon cancer cells, depending on their metastatic potentials. Normal cells of two origins had characteristically very low levels of GnT-III and -V activities, whereas hepatoma and colon cancer cells contained high levels of activities. These data were supported by RT-PCR and Northern blot analyses, showing that the expression of GnT-III and -V mRNAs were increased in proportion to the enzymatic activities. The increased GnT-III, md -V activities were also correlated with increased glycosylation of the cellular glycoproteins in hepatoma and colon cancer cells, as examined by lectin blotting analysis by using wheat germ glutinin (WGA), erythroagglutinating phytohemagglutinin (E-PHA), leukoagglutinating phytohemagglutinin (L-PHA), and concanavalin A (Con A). Treatment with retinoic acid, a differentiation agent, resulted in decreases of both GnT-III and -V activities of HepG2 and HepG3 cells. In colon carcinoma cells, however, treatment with retinoic acid resulted in a reduction of GnT-V activity, but not with GnT-III activity. Although the mechanism underlying the induction of these mzymes is unclear, oligosaccharides in many glycoproteins have been observed of cancer cells.

Keywords

References

  1. Alverez, K., C. Haswell, M. St. Clair, G. S. Pemg, M. Shorebah, M. Pierce, and N. Fregien. 2002. Sequences of the mouse N-acetylglucosaminyltransferase V (Mgat5) mRNA and an mRNA expressed by an Mgatfi-deficient cell line. Glycobiology 12: 389- 394.
  2. Arango, J. and M. Pierce. 1988. Comparison of Nacetylglucosaminyltransferase V activities in Rouse sarcomatransformed baby hamster kidney (RS-BHK) and BHK cells. J. Cell. Biochem. 37: 225- 231.
  3. Brockhausen, I., R. A. Romero, and A. Herscovics. 1991. Glycosyltransferase changes upon differentiation of CaCo-2 human colonic adenocarcinoma cells. Cancer Res. 51: 3136-3142.
  4. Chung, T. W, S. K. Moon, Y C. Lee, J. G. Kim, J. H. Ko, and C. H. Kim. 2002. Enhanced expression of matrix metalloproteinase-S (MMP-9) by hepatitis B virus infection into liver cells. Arch. Biochem. Biophys. 408: 147- 154.
  5. Chung, T. W, Y C. Lee, J. H. Ko, and C. H. Kim. 2003. Hepatitis B virus X protein (HBx) is a negative regulator of PTEN in liver cells: Involvement of AKT activation. Cancer Res. 63: 3453- 3458.
  6. Chung, Y. J., J. R. Jeong, B. C. Lee, J. Y Kim, Y I. Park, and J. Y Ro. 2003. Phospholipase D in guinea pig lung tissue membrane is regulated by cytosolic ARF proteins. J. Microbiol. Biotechnol. 13: 897- 905.
  7. Dennis, J. W. and S. Kaferte. 1989. Oncodevelopmental expression of GlcNAc $\beta$1-6Manl-branched asparaginelinked oligosaccharides in murine tissues and human breast carcinomas. Cancer Res. 49: 945- 960.
  8. Dennis, J. W, S. Laferte, C. Waghorne, M. L. Breitman, and R. S. Kerbel. 1987. $\beta$-1,6 Branching of Asn-linked oligosaccharides is directly associated with metastasis. Science 236: 582- 585.
  9. Fournet, B.,J. Motreuil, G. Strecker, L. Dorland, J. Haverkamp, F. G. Vilegenthart, J. P. Binette, and K. Schmid. 1978. Determination of the primary structures of 16 asialocarbohydrate units derived from human plasma alpha l-acid glycoprotein by 360-MHZ IH NMR spectroscopy and permethylation analysis. Biochemistry 17: 5206- 5214.
  10. Guo, H. B., A. L. Jiang, T. Z. Ju, and H. L. Chen. 2000. Opposing changes in Nsacetylglucosaminyltransferase-V and -III during the cell cycle and all-trans retinoic acid treatment of hepatocarcinoma cell line. Biochim. Biophys. Acta 1495: 297- 307.
  11. Guo, H. B., O. S. Zhang, and H. L. Chen. 2000. Effects of H-ras and v-sis overexpression on N-acetylglucosaminyltransferase V and metastasis-related phenotypes in human hepatocarcinoma cells. J. Cancer Res. Clin. Oncol. 126: 263- 270.
  12. Gu, J., A. Nishikawa, N. Tsuruoka, M. Ohno, N. Yamaguchi, K. Kanagawa, and N. Taniguchi. 1993. Purification and characterization of UDP-N-GIcNAc:f3-6-D-mannoside 131,6- N-acetylglucosaminyltransferase III from a human lung cancer cell line. J. Biochem. 113: 614- 619.
  13. Hakomori, S. 1989. Aberrant glycosylation in tumors and tumor-associated carbohydrate antigens. Adv. Cancer Res. 52: 257- 331.
  14. Hase, S., Y Ibuki, and T. Ikenaka. 1984. Reexamination of the pyridylamination used for fluorescence labeling of oligo saccharides and its application to glycoproteins. J. Biochem. 95: 197- 203.
  15. Jun, D. Y, S. W Rue, B. W Kim, and Y H. Kim. 2003. Detection of mitotic centromere-associated kinesin (MCAK) during cell-cycle progression of human Jurkat T cells using polyclonal antibody raised against its N-terrninal region overexpressed in E. coli. J. Microbiol. Biotechnol. 13: 912-919.
  16. Kim, C. H. 2004. Increased expression of Nacetylglucosarninyltransferase- V in human hepatoma cells by retinoic acid and 1alpha,25-dihydroxyvitamin D3. Int. J. Biochem. Cell Bioi. 36: 2307- 2319. https://doi.org/10.1016/j.biocel.2004.04.021
  17. Kim, S. W, S. H. Lee, K. S. Kim, C. H. Kim, Y. K. Choo, and Y C. Lee. 2002. Isolation and characterization of the promoter region of the human GM3 synthase gene. Biochim. Biophys. Acta 1578: 84- 99.
  18. Kim, C. H., Y. J. Kim, B. S. Kang, and Y. C. Lee. 1997. Sequence of 5'-flanking region of the human GnT-III gene and secretion inhibition of hepatic viral coat-proteins by introduction of the gene in Hep3B. International Symposium on Glycosyltransferases and Cellular Communications, p. 106.
  19. Kim, K. S., C. H. Kim, D. Y. Shin, and Y C. Lee. 1997. Molecular cloning and expression of cDNAs encoding mouse Gal 1,3(4)GaINAc 2,3sialyltransferase (mST3Gal III) and Gal 1,4(3)GaINAc 2,3sialyltransferase (mST3Gal IV). J. Biochem. Mol. BioI. 30: 95- 100.
  20. Kim, K S., C. H. Kim, J. K Kim, and Y. C. Lee. 2000. Inhibitory effects of oriental medicinal herbs on enzymatic activity of sialyltransferases (ST3GalI and ST6GalI) associated with cancer. J. Life Sci. 10: 51- 56.
  21. Kim, K W., K S. Kim, C. H. Kim, J. K Kim, and Y. C. Lee. 1999. Molecular cloning and sequence analysis of human GM3 synthase (hST3Gal V). J. Biochem. Mol. Biol. 32: 409-413.
  22. Kim, K w., S. W. Kim, K S. Min, C. H. Kim, and Y. C. Lee. 200I. Genomic structure of human GM3 synthase gene (hST3Gal V) and identification of mRNA isoforms in the 5'untranslated region. Gene 273: 163- 171.
  23. Kim, S. W., N. Y. Kang, S. H. Lee, K W. Kim, K S. Kim, J. H. Lee, C. H. Kim, and Y. C. Lee. 2003. Genomic structure and promoter analysis of human NeuAcalpha2,3Gal beta I,3GaINAcalpha2,6-sialyltransferase (hST6GaINAc IV) gene. Gene 305: 113- 120.
  24. Kim, S. W., S. H. Lee, K. S. Kim, C. H. Kim, Y. K Chao, and Y. C. Lee. 2002. Isolation and characterization of the promoter region of the human GM3 synthase gene. Biochim. Biophys. Acta 1578: 84- 89.
  25. Kim, Y. J., K S. Kim, C. H. Kim, and Y. C. Lee. 1996. Molecular cloning and expression of human Gall ,3GaINAc 2,3sialyltransferase (hST3GalII). Biochem. Biophys. Res. Comm. 228: 324- 327.
  26. Kim, Y. J., K S. Kim, S. I. Do, C. H. Kim, S. K Kim, and Y.C. Lee. 1997. Molecular cloning and expression of human 2,8sialyltransferase (hST8Sia V). Biochem. Biophys. Res. Comm. 235: 327- 330.
  27. Kim, Y. J., K S. Kim, T. W. Chung, I. S. Choe, Y. C. Lee, and C. H. Kim. 1996. Sequence analysis of the 5'-flanking region of the gene encoding human N-acetylglucosaminyltransferase III. Gene 170: 281- 283.
  28. Kobata, A. and K. Yamashita. 1984. Structural change of sugar chains of glycoproteins by cell transformation and its application to the diagnosis of cancer. Gan No Rinsho 30: 545- 551 (In Japanese).
  29. Koenderman, A H. L., P. L. Koppen, C. A M. Koeleman, and D. H. Van den Eijnden. 1989. N-acetylglucosarninyltransferase III, IV, V in Novikoff ascites tumor cells, mouse lymphoma cells and hen oviduct. Eur. J. Biochem. 181: 651- 655.
  30. Lee, D. H. 2002. Determination of optimum aggregates of porcine hepatocytes as a cell source of a bioartificial liver. J. Microbiol. Biotechnol. 12: 735- 739.
  31. Lee, J. w., E. Y. Song, T. W.Chung, K S. Kim, T. H. Chung, Y. I. Yeorn, and C. H. Kim. 2004. Hyperexpression of N-acetylglucosaminyltransferase-III in liver tissues of transgenic mice causes fatty body and obesity through severe accumulation of Apo-AI and ApoB. Arch. Biochem. Biophys.426b 18-31.
  32. Lee, Y. c, Y. J. Kim, K S. Kim, H. N. Kim, S. I. Do, and C. H. Kim. 1988. Cloning and expression of cDNA for a Human Sia 2,3Gal 1,4GaINA: 2,8Sialyltransferase (hST8Sialll). Arch. Biochem. Biophys. 360: 41- 46.
  33. Lowry, O. H., N. H. Rosenbrough, A L. FaIT, and H. Randall. 1951. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193: 265- 275.
  34. Miyoshi, E., A Nishikawa, Y. Ihara, J. Gu, T. Sugiyama, N. Hayashi, H. Fusamoto, T. Kamada, and N. Taniguchi. 1993. N-acetylglucosaminyltransferase III and V messenger RNA levels in LEC rats during hepatocarcinogenesis. Cancer Res. 53: 3899- 3902.
  35. Miyoshi, E., Y. lhara, A. Nishikawa, H. Saito, N. Uozumi, N. Hayashi, H. Fusamoto, K Takenobu, and N. Taniguchi. 1995. Gene expression of N-acetylglucosaminyltransferase III and V: A possible implication for liver regeneration. Hepatology 22: 1847- 1855.
  36. Mizoguchi, A, S. Takasaki, S. Maeda, and A Kobata. 1984. Changes in asparagine-linked sugar chains of human promyelocytic leukemic cells (HL-60) during monocytoid differentiation and myeloid differentiation. Appearance of high mannose-type oligosaccharides in neutral fraction. J. Biol. Chem. 259: 11943- 11948.
  37. Nishikawa, A., S. Fujii, T. Sugiyama, N. Hayashi, and N. Taniguchi. 1988. High expression of Nacetylglucosaminyltransferase III in 3'-methyl DAB-induced hepatoma and ascites hepatoma. Biochem. Biophys. Res. Comm. 152: 107- 112.
  38. Ohno, M., A. Nishikawa, M. Koketsu, H. Taga, Y. Endo, T. Hada, K Higashino, and N. Taniguchi. 1992. Enzymatic basis of sugar structures of alpha-fetoprotein in hepatoma and hepatoblastoma cell lines: Correlation with activities of alpha 1- 6 fucosyltransferase and Nacetylglucosaminyltransferases III and V. Int. J. Cancer 51: 315-317.
  39. Park, C., U. H. Jin, Y. C. Lee, T. J. Cho, and C. H. Kim. 1999. Characterization of UDP-N-acetylglucosamine:$\alpha$-6-D-mannoside $\beta$-I ,6-N-acetylglucosaminyltransferase-V from human hepatoma cell line Hep3B. Arch. Biochem. Biophys. 367: 281- 288.
  40. Pierce, M. and J. Arango. 1986. Rouse sarcoma-transformed baby hamster kidney cells express higher levels of asparaginelinked tri- and tetraantennary glycopeptides containing GlcNAc$\beta$(I,6)Man-$\beta$( I,6)Man and poly-N-acetyllactosamine sequences than baby hamster kidney cells. J. Biol. Chem. 261: 10772- 10777.
  41. Schachter, H. 1986. Biosynthetic controls that determine the branching and micro heterogeneity of protein-bound oligosaccharides. Biochem. Cell. Bioi. 64: 163- 181.
  42. Shim, J. K, D. W. Kim, T. W. Chung, J. K Kim, J. I. Suh, C. Park, Y. C. Lee, T. H. Chung, E. Y. Song, and C. H. Kim. 2001. Characterization of a cell line HFH-T2, producing viral particles, from primary human fetal hepatocytes infected with hepatitis B virus. J. Microbiol. Biotechnol. 11: 186-192.
  43. Song, E. Y., K A. Kim, Y. D. Kim, D. H. Kwon, H. S. Lee, T. W. Chung, S. M. Byun, and C. H. Kim. 2000. A simplified procedure for the purification of human Nacetylglucosaminyltransferase- Ill from human hepatocellular carcinoma tissues. Biotechnol. Lett. 22: 201- 204.
  44. Song, E. Y., S. G. Kang, Y. C. Lee, Y. G. Park, T. H. Chung, D. H. Kwon, S. M. Byun, and C. H. Kim. 2001. Expression of bisecting N-acetylglucosaminyltransferase-Ill in human hepatocarcinoma tissues, fetal liver tissues, and hepatoma cell lines of Hep3E and HepG2. Cancer Invest. 19: 797-805.
  45. Stanley, P. 2002. Biological consequences of overexpressing or eliminating N-acetylglucosaminyltransferase-III in the mouse. Biochim. Biophys. Acta 1573: 363- 368.
  46. Yamashita, K., A. Hitoi, and A. Kobata. 1983. Structural determinants of Phaseolus vulgaris erythroagglutinating lectin for oligosaccharides. J. Biol. Chem. 258: 14753- 14755.
  47. Yamashita, K., A. Hitoi,N. Taniguchi, N. Yokosawa, Y Tsukada, and A. Kobata. 1983. Enzymatic basis for the structure changes of asparagine-linked sugar chains of membrane glycoproteins of baby hamster kidney cells induced by polyoma transformation. Cancer Res. 43: 5059- 5063.