Regular Article

British Journal of Cancer (2001) 84, 1556–1563. doi:10.1054/bjoc.2001.1815 www.bjcancer.com
Published online 29 May 2001

Regulation of differentiation- and proliferation-inducers on Lewis antigens, alpha-fucosyltransferase and metastatic potential in hepatocarcinoma cells

F Liu1,*, H-L Qi1,* and H-L Chen1

1Key Laboratory of Glycoconjugate Research, Ministry of Health, Department of Biochemistry, School of Medicine, Fu-Dan University, Shanghai, 200032, China

*These two authors contributed equally to this manuscript.

Received 11 September 2000; Revised 1 February 2001; Accepted 9 February 2001.

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Abstract

The expressions of Lewis (Le) antigens, alpha-1,3/1,4 fucosyltransferases (alpha-1,3/1,4 FuTs), and metastatic potential after the treatment of 2 differentiation inducers, all-trans retinoic acid (ATRA), 8-bromo-cyclic 3',5'adenosine monophosphate (8-Br-cAMP); and 2 proliferation inducers, epidermal growth factor (EGF) and phobol-12-myristate-13-acetate (PMA), on 7721 human hepatocarcinoma cell line were studied. Cell adhesion to human umbilical vein endothelial cells (HUVEC), cell migration through transwell and invasion through matrigel were selected as the indexes of metastatic potential-related phenotypes. Using fluorescence-labelled antibodies and flow-cytometric analysis, it was found that 7721 cells mainly expressed sialyl Lewis X (SLex) and a less amount of sialyl dimeric Lewis X (SDLex) antigens on the cell surface. Their expressions were down-regulated by ATRA, and up-regulated by EGF. SLex antigen was also decreased and increased by the treatment of 8-Br-cAMP and PMA respectively. With Northern blot to detect the mRNAs of alpha-1,3/1,4 FuTs, the main enzymatic basis for the change in SLex expression was found to be the alteration of the expression of alpha-1,3 FuT-VII. It was evidenced by the observations that alpha-1,3 FuT-VII was the main alpha-1,3/1,4 FuT in 7721 cells, while alpha-1,3/1,4 FuT-III and alpha-1,3 FuT-VI were expressed rather low. The changes in the expressions of SLex antigen and alpha-1,3 FuT-VII resulted in the altered cell adhesion to tumour necrosis factor-alpha stimulated HUVEC, since only the monoclonal antibody of the SLex, but not other monoclonal antibodies blocked the adhesion of 7721 cells to HUVEC. The migration and invasion of 7721 cells were also reduced by the treatment of ATRA or 8-Br-cAMP, and elevated by EGF or PMA. The above findings indicate that the metastatic potential of 7721 cells is suppressed by differentiation-inducers and promoted by proliferation-inducers. © 2001 Cancer Research Campaign http://www.bjcancer.com

Keywords:

human hepatocarcinoma cells, differentiation-inducer, proliferation-inducer, Lewis antigen, alpha-fucosyltransferase, metastasis-related phenotype

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References

  1. Ai ZW, Zha XL, Liu Y and Chen HL (1990) Effects of retinoic acid on hepatocarcinoma cell line. J Tumor Mark Oncol 5: 59–70
  2. Ai ZW, Zha XL, Tang H and Chen HL (1991a) Reversing effect of retinoic acid on some phenotypes of human hepatocarcinoma cell line. Chin J Oncol 13: 9–11
  3. Ai ZW, Chen HL, Gu JR, Jiang HO and Long J (1991b) Inhibition by retinoic acid of the expression of the oncogene ras in human hepatocarcinoma cell line. Acta Biochim Biophy Sinica 23: 453–457
  4. Chai XY, Qin H and Chen HL (1993a) Protein kinase and cell differentiation, inhibition of tyrosine protein kinase by two inducers of differentiation. Chin Biochem J 9: 162–167
  5. Chai XY, Shen YF and Chen HL (1993b) Effect of phorbor ester on protein kinase C and tyrosine protein kinase in human hepatocarcinoma cell line. Chin J Oncol 15: 182–184
  6. Chai XY and Chen HL (1994a) The effects of two differentiation-inducers on serine-protein kinase activity in human hepatocarcinoma cell line. Acta Acad Med Shanghai 21: 170–175
  7. Chai XY and Chen HL (1994b) Expression of oncogenes during induced differentiation of human hepatocarcinoma cell line. Chin J Cancer Res 6: 3–8
  8. Chai XY, Chen HL, Zhou XM, Qian LF, Chen SH, Jiang HO and Gu JR (1994c) The opposite effects of retinoic acid and phorbol ester on the expression of c-myc and IGF-II genes in human hepatocarcinoma cells. Acta Oncol Sinica 4: 229–231
  9. Chen HL, Dong SC, Ju TZ and Yang XP (1995) Effect of retinoic acid on the structure of N-glycan on the surface of human hepatocarcinoma cells and its enzymatic Mechanism. J Cancer Res Clin Oncol 121: 397–401
  10. Dong SC, Yang XP and Chen HL (1994) Effect of dibutyryl cAMP on the type and antennary number of N-glycans on the surface of human hepatocarcinoma cell line SMMC 7721. Acta Biochim Biophys Sinica 26: 43–49
  11. Hakomori SI (1996) Tumor malignancy defined by aberrant glycosylation and sphingo(glyco)lipid metabolism. Cancer Res 56: 5309–5318 | PubMed | ISI | ChemPort |
  12. Issiki S, Togayachi A, Kudo T, Nishihara S, Watanabe M, Kobata T, Kitajima M, Shiraishi H, Sasaki K, Andoh T and Narimatsu H (1999) Cloning, expression, and charisterazation of novel UDP-galactose: beta-N-actylglucosamine beta1,3-galactosyltransferase (beta3Gal-T5) responsible for synthesis of type I chain in colorectal and pancreatic epithelia and tumor cells derived therefrom. J Biol Chem 274: 12499–12507 | PubMed | ChemPort |
  13. Ito H, Hiraiwa N, Sawada-Kasugai M, Akamatsu S, Tachikawa T, Kasai Y, Akiyama S, Ito K, Takagi H and Kannagi R (1997) Altered mRNA expression of specific molecular species of fucosyl-and sialyl-transferases in human colorectal cancer tissues. Int J Cancer 71: 5560–5564
  14. Jorgensen T, Berner A, Kaalhus O, Tveter KJ, Danielsen HE and Bryne M (1995) Up-regulation of the oligosaccharide sialyl Lewis X: a new prognosis parameter in metastatic prostate cancer. Cancer Res 55: 1817–1819 | PubMed | ChemPort |
  15. Kansas GS. Selectins and their ligands: (1996) Current concepts and controversies. Blood 88: 3259–3287 | PubMed | ISI | ChemPort |
  16. Kudo K, Ikehara Y, Togayachi A, Kaneko M, Hiraga T, Sasaki K and Narimatsu H (1998) Expression, cloning and characterization of a novel murine alpha1,3-fucosyltransferase, mFuc-IX, that synthesis the Lewis X (CD15) epitope in brain and kidney. J Biol Chem 273: 26729–26738
  17. Kudo T, Ikehara Y, Togayachi A, Morozumi K, Watanabe M, Nakamura S and Narimatsu H (1998) Up-regulation of a set of glycosyltransferase gene in human colorectal cancer. Lab Invest 78: 797–811 | PubMed | ISI | ChemPort |
  18. Kukowska-Latallo JF, Larsen RD, Nair RP and Lowe JB (1990) A cloned human cDNA determines expression of a mouse stage-specific embryonic antigen and the Lewis blood group (alpha1,3/1,4) fucosyltransferase. Genes Develop 4: 1288–1303
  19. Liu F, Qi HL and Chen HL (2000) Effects of all- trans retinoic acid and epidermal growth factor on the expression of nm23 -H1 in human hepatocarcinoma cells. J Cancer Res Clin Oncol 126: 85–90 | PubMed | ISI | ChemPort |
  20. Lowe JB, Kukowska-Latallo JF, Nair RP, Larsen RD, Marks RM, Marcher BA, Kelly RJ and Ernst LK (1991) 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
  21. Nakamori S, Kameyama M, Imaoka S, Furukawa H, Ishikawa O, Sasaki T, Kabuto T, Iwanaga T, Matsushita Y and Irimura Y (1993) Increased expression of sialyl Lewis X antigen correlates with poor survival in patients with colorectal carcinoma: clinico-pathological and immunohisto-chemical study. Cancer Res 53: 3632–3637 | PubMed | ISI | ChemPort |
  22. Narimatsu H (1998) Human fucosyltransferases, Tissue distribution of blood group antigens, cancer associated antigens and fucosyltransferase. Protein Nucleic Acid Enzyme 48: 2394–2403
  23. Nicholas SF and Paolo SC (1996) Transcription factors coupled to the cAMP signalling pathway. Biochim Biophys Acta 1288: F101–F121 | Article | PubMed | ISI | ChemPort |
  24. Nishizuka Y (1992) Intracellular signaling by hydrolysis of phospholipids and activation of protein kinase C. Science 258: 607–614 | Article | PubMed | ISI | ChemPort |
  25. Ogawa J, Tsurumi T, Yamada S, Koide S and Shohtsu A (1994) Blood vessel invasion and expression of sialyl Lewis X and proliferating cell nuclear antigen in non-small cell lung cancer. Cancer 73: 1177–1183
  26. Ogawa J, Inoue H and Koide S (1997) alpha-2,3-sialyltransferase type 3N and alpha-1,3-fucosyl-transferase type VII are related to sialyl Lewis X synthesis and patient survival from lung carcinoma. Cancer 79: 1678–1685
  27. Sagestrom CG and Sive HL (1996) RNA blot analysis, In A Laboratory Guide to RNA: Isolation, Analysis, and Synthesis. Krieg PA (ed) pp 83–103. Wiley-Liss Inc. London, New York, Tokyo
  28. Sasaki K, Kurata K, Funayama K, Nagata M, Watanabe E, Ohta S, Hanat N and Nishi T (1994) Expression cloning of a novel alpha1,3-fucosyltransferase that is involved in biosynthesis of the sialyl Lewis X carbohydrate determinants in leukocytes. J Biol Chem 269: 14730–14737 | PubMed | ISI | ChemPort |
  29. Shen YF, Chai XY and Chen HL (1993) Effect of phorbol ester on tyrosine protein kinase in human hepatocarcinoma cell line. Acta Biochim Biophy Sinica 25: 537–540
  30. Takada A, Ohmori K, Yoneda T, Tsuyuoka K, Hasegawa K, Kiso M and Kannagi R (1993) Contribution of carbohydrate antigens sialyl Lewis A and sialyl Lewis X to adhesion of human cancer cells to vascular endothelium. Cancer Res 53: 354–361 | PubMed | ISI | ChemPort |
  31. Taniguchi N, Miyoshi E, Ko JH, Ikeda Y and Ihara Y (1999) Implication of N-acetylglucosaminyl transferases III and V in cancer: gene regulation and signaling mechanism. Biochim Biophys Acta 1455: 287–300 | PubMed |
  32. van der Geer P, Hunter T and Lindberg RA (1994) Receptor protein-tyrosine kinase and their signal transduction pathways. Ann Rev Cell Biol 10: 251–337 | Article | PubMed | ChemPort |
  33. Welch DR (1997) Technical considerations for studying cancer metastasis in vivo. Clin Exp Metastasis 15: 272–306 | Article | PubMed | ISI | ChemPort |
  34. Weston BW, Nair RP, Jarsen RD and Lowe JB (1992a) Isolation of a novel human alpha1,3 fucosyl-transferase gene and molecular comparison to the human Lewis blood group alpha(1,3/1,4) fucosyltransferase gene, synthetic, homologous, nonallelic genes encoding enzymes with distinct acceptor substrate specificity. J Biol Chem 267: 4152–4160 | PubMed | ChemPort |
  35. Weston BW, Smith PL, Kelly RJ and Lowe JB (1992b) Molecular cloning of a fourth member of a human alpha(1,3) fucosyltransferase gene family. Multiple homologous sequences that determine expression of the Lewis X, sialyl Lewis X, and difucosyl sialyl Lewis X antigens. J Biol Chem 267: 24575–24584
  36. Xia BY, Guo HB and Chen HL (1998) Dual regulation of tyrosine protein kinase and protein kinase C on N-acetylglucosaminyltransferase V. Acta Biochim Biophs Sinica 30: 427–431
  37. Yu D, Wang SS, Dulki KM, Tsai CM, Nicolson GL and Hung MC (1994) C-erbB2/neu overexpression enhances metastatic potencial of human lung cancer cells by induction of metastasis-associated properties. Cancer Res 54: 3260–3266 | PubMed | ISI | ChemPort |