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Japanese Journal of Cancer Research : Gann logoLink to Japanese Journal of Cancer Research : Gann
. 1990 Dec;81(12):1286–1292. doi: 10.1111/j.1349-7006.1990.tb02692.x

Tumor‐promoting Phorbol Ester Induces Alterations of Sialidase and Sialyltransferase Activities of JB6 Cells

Taeko Miyagi 1,, Junji Sagawa 1, Toshio Kuroki 2, Yutaka Matsuya 3,, Shigeru Tsuiki 1,
PMCID: PMC5918020  PMID: 2125997

Abstract

Sialidase and sialyltransferase activities were studied in JB6 mouse epidermal cells before and after exposure to phorbol ester, 12‐O‐tetradecanoyl phorbol‐13‐acetate (TPA), which irreversibly induces anchorage‐independent growth and tumorigenicity. JB6 cells exhibited sialidase activities toward 4‐methylumbelliferyl‐α‐d‐N‐acetylneuraminic acid (4MU‐NeuAc) and gangliosides at pH 4.5 in the particulate fraction but apparently not in the cytosol at pH 4.5 or 6.0. In JB6 cells exposed to TPA and in the anchorage‐independent transformants, the sialidase activity toward 4MU‐NeuAc was decreased and the activity toward gangliosides was increased compared with those in untreated JB6 cells. Immunological analysis with antisera against membrane‐associated sialidases I and II revealed that plasma membrane‐associated sialidase I was increased and lysosomal membrane‐associated sialidase II was decreased under these conditions. TPA treatment also affected the sialyltransferase activities of JB6 cells: an elevation of the transfer activities toward asialo‐orosomucoid and asialo‐porcine submaxillary mucin but a reduction of GM3 and GD3 synthase activities were observed on exposure to TPA and in cells transformed by TPA to retain anchorage‐independency. These results suggest that an increase in sialic acid bound to glycoproteins and a decrease in that bound to glycolipids may occur in JB6 cells exposed to TPA and in the anchorage‐independent transformants.

Keywords: Sialidase, JB6 cell, Phorbol ester, Anchorage independence, Ganglioside

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REFERENCES

  • 1. ) Warren , L. , Buck , C. A. and Tuszynski , G. P.Glycopeptide changes and malignant transformation, a possible role for carbohydrate in malignant behaviour . Biochim. Biophys. Acta , 516 , 97 – 127 ( 1978. ). [DOI] [PubMed] [Google Scholar]
  • 2. ) Hakomori , S.Aberrant glycosylation in tumor and tumor‐associated carbohydrate antigens . Adv. Cancer Res. , 52 , 257 – 331 ( 1989. ). [DOI] [PubMed] [Google Scholar]
  • 3. ) Reutter , W. , Kottgen , E. , Bauer , C. and Gerok , W.Biological significance of sialic acid . In “ Sialic Acid: Chemistry, Metabolism and Functions ,” ed. Schauer R. , pp. 263 – 305 ( 1982. ). Springer‐Verlag; , Vienna, New York . [Google Scholar]
  • 4. ) Symington , B. E. , Symington , F. W. and Rohrschneider , L. R.Phorbol ester induces increased expression, altered glycosylation, and reduced adhesion of K562 erythroleukemia cell fibronectin receptors . J. Biol. Chem. , 264 , 13258 – 13266 ( 1989. ). [PubMed] [Google Scholar]
  • 5. ) Yogeeswaran , G. and Salk , P.Metastatic potential is positively correlated with cell surface sialylation of cultured murine tumor cell lines . Science , 212 , 1514 – 1516 ( 1981. ). [DOI] [PubMed] [Google Scholar]
  • 6. ) Fogel , M. , Altevogt , P. and Schirrmacher , V.Metastatic potential severely altered by changes in tumor cell adhesiveness and cell‐surface sialylation . J. Exp. Med. , 157 , 371 – 376 ( 1983. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 7. ) Collard , J. G. , Schijven , J. F. , Bikker , A. , La Riviere , G. , Bolscher , J. G. M. and Roos , E.Cell surface sialic acid and the invasive and metastatic potential of T‐cell hybridomas . Cancer Res. , 46 , 3521 – 3527 ( 1986. ). [PubMed] [Google Scholar]
  • 8. ) Miyagi , T. and Tsuiki , S.Rat‐liver lysosomal sialidase. Solubilization, substrate specificity and comparison with the cytosolic sialidase . Eur. J. Biochem. , 141 , 75 – 81 ( 1984. ). [DOI] [PubMed] [Google Scholar]
  • 9. ) Miyagi , T. and Tsuiki , S.Purification and characterization of cytosolic sialidase from rat liver . J. Biol. Chem. , 260 , 6710 – 6716 ( 1985. ). [PubMed] [Google Scholar]
  • 10. ) Miyagi , T. and Tsuiki , S.Evidence for sialidase hydrolyzing gangliosides GM2 and GM1 in rat liver plasma membrane . FEBS Lett. , 206 , 223 – 228 ( 1986. ). [DOI] [PubMed] [Google Scholar]
  • 11. ) Miyagi , T. , Sagawa , J. , Konno , K. , Handa , S. and Tsuiki , S.Biochemical and immunological studies on two distinct ganglioside‐hydrolyzing sialidases from the particulate fraction of rat brain . J. Biochem. , 107 , 787 – 793 ( 1990. ). [DOI] [PubMed] [Google Scholar]
  • 12. ) Miyagi , T. , Sagawa , J. , Konno , K. and Tsuiki , S.Immunological discrimination of intralysosomal, cytosolic, and two membrane sialidases present in rat tissues . J. Biochem. , 107 , 794 – 798 ( 1990. ). [DOI] [PubMed] [Google Scholar]
  • 13. ) Miyagi , T. , Goto , T. and Tsuiki , S.Sialidase of rat hepatomas: qualitative and quantitative comparison with rat liver sialidase . Gann , 75 , 1076 – 1082 ( 1984. ). [PubMed] [Google Scholar]
  • 14. ) Sagawa , J. , Miyagi , T. and Tsuiki , S.Membrane‐associated sialidase of rat liver and its decrease in hepatomas . Jpn. J. Cancer Res. , 79 , 69 – 73 ( 1988. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 15. ) Miyagi , T. , Konno , K. , Sagawa , J. and Tsuiki , S.Neoplastic alteration of a membrane‐associated sialidase of rat liver . Jpn. J. Cancer Res. , 81 , 915 – 919 ( 1990. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 16. ) Miyagi , T. and Tsuiki , S.Purification and characterization of β‐galactoside (α2→6) sialyltransferase from rat liver and hepatomas . Eur. J. Biochem. , 126 , 253 – 261 ( 1982. ). [PubMed] [Google Scholar]
  • 17. ) Miyagi , T. , Koseki , M. and Tsuiki , S.Comparative study of the levels of sialyltransferases responsible for the formation of sugar chains in glycoproteins and gangliosides in rat liver and hepatomas . Jpn. J. Cancer Res. , 79 , 742 – 749 ( 1988. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 18. ) Colburn , N. H. , Bruegge , W. F. V. , Bates , J. R. , Gray , R. H. , Rossen , J. D. , Kelsey , W. H. and Shimada , T.Correlation of anchorage‐independent growth with tumorigenicity of chemically transformed mouse epidermal cells . Cancer Res. , 38 , 624 – 634 ( 1978. ). [PubMed] [Google Scholar]
  • 19. ) Colburn , N. H. , Former , B. F. , Nelson , K. A. and Yuspa , S. H.Tumour promoter induces anchorage independence irreversibly . Nature , 281 , 589 – 591 ( 1979. ). [DOI] [PubMed] [Google Scholar]
  • 20. ) Srinivas , L. , Gindhart , T. D. and Colburn , N. H.Tumor‐promoter‐resistant cells lack trisialoganglioside response . Proc. Natl. Acad. Sci. USA , 79 , 4988 – 4991 ( 1982. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 21. ) Tsuiki , S. , Hashimoto , Y. and Pigman , W.Comparison of procedures for the isolation of bovine submaxillary mucin . J. Biol. Chem. , 236 , 2172 – 2178 ( 1961. ). [PubMed] [Google Scholar]
  • 22. ) de Salegui , M. and Plonska , H.Preparation and properties of porcine submaxillary mucins . Arch. Biochem. Biophys. , 129 , 49 – 56 ( 1969. ). [DOI] [PubMed] [Google Scholar]
  • 23. ) Hosoi , J. , Abe , E. , Suda , T. , Colburn , N. H. and Kuroki , T.Induction of anchorage‐independent growth of JB6 mouse epidermal cells by 12,25‐dihydroxyvitamin D3 . Cancer Res. , 46 , 5582 – 5586 ( 1986. ). [PubMed] [Google Scholar]
  • 24. ) Lowry , O. H. , Rosebrough , N. J. , Farr , A. L. and Randall , R. J.Protein measurement with the Folin phenol reagent . J. Biol. Chem. , 193 , 265 – 275 ( 1951. ). [PubMed] [Google Scholar]
  • 25. ) Warren , L.The thiobarbituric acid assay of sialic acids . J. Biol. Chem. , 234 , 1971 – 1975 ( 1959. ). [PubMed] [Google Scholar]
  • 26. ) Chien , J‐L. , Williams , T. and Basu , S.Biosynthesis of a globoside‐type glycosphingolipid by a β‐N‐acetyl‐galactosaminyltransferase from embryonic chicken brain . J. Biol. Chem. , 248 , 1778 – 1785 ( 1973. ). [PubMed] [Google Scholar]
  • 27. ) Yogeeswaran , G. and Hakomori , S.Cell contact‐dependent ganglioside changes in mouse 3T3 fibroblasts and a suppressed sialidase activity on cell contact . Biochemistry , 14 , 2151 – 2156 ( 1975. ). [DOI] [PubMed] [Google Scholar]
  • 28. ) Schengrund , C. L. , Lausch , R. N. and Rosenberg , A.Sialidase activity in transformed cells . J. Biol. Chem. , 248 , 4424 – 4428 ( 1973. ). [PubMed] [Google Scholar]
  • 29. ) Yamashita , K. , Ohkura , T. , Tachibana , Y. , Takasaki , S. and Kobata , A.Comparative study of the oligosaccharides released from baby hamster kidney cells and their polyoma transformant by hydrazinolysis . J. Biol. Chem. , 259 , 10834 – 10840 ( 1984. ). [PubMed] [Google Scholar]
  • 30. ) Collard , J. G. , van Beek , W. P. , Janssen , J. W. G. and Shijven , J. F.Transfection by human oncogenes: concomitant induction of tumorigenicity and tumor‐associated membrane alterations . Int. J. Cancer , 35 , 207 – 214 ( 1985. ). [DOI] [PubMed] [Google Scholar]
  • 31. ) Pierce , M. and Arango , J.Rous sarcoma virus‐transformed baby hamster kidney cells express higher levels of asparagine‐linked tri‐ and tetraantennary glycopeptides containing [GlcNAc‐β(1,6)Man‐α(1,6)] and poly‐N‐acetyllactosamine sequences than baby hamster kidney cells . J. Biol. Chem. , 261 , 10772 – 10777 ( 1986. ). [PubMed] [Google Scholar]
  • 32. ) Santer , U. V. , De Santis , R. , Hard , K. J. , van Kuik , J. A. , Vliegenthart , J. F. G. , Won , B. and Glick , M. C.N‐Linked oligosaccharide changes with oncogenic transformation require sialylation of multiantennae . Eur. J. Biochem. , 181 , 249 – 260 ( 1989. ). [DOI] [PubMed] [Google Scholar]
  • 33. ) Momoi , T. , Shinmoto , M. , Kasuya , J. , Senoo , H. and Suzuki , Y.Activation of CMP‐N‐acetylneuraminic acid: lactosylceramide sialyltransferase during the differentiation of HL‐60 cells induced by 12‐O‐tetradecanoylphorbol‐13‐acetate . J. Biol. Chem. , 261 , 16270 – 16273 ( 1986. ). [PubMed] [Google Scholar]

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