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Expression of α2,8-sialyltransferase (GD3 synthase) gene in human cancer cell lines: high level expression in melanomas and up-regulation in activated T lymphocytes

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Abstract

GD3 Synthase (α2,8sialyltransferase) (EC 2.4.99.8) cDNA has been cloned by eukaryotic cell expression cloning. Using this cDNA as a probe, the expression level of the gene in human cancer cell lines was analysed by Northern blotting and RT-PCR, then correlated with the ganglioside expression and enzyme activity. Melanoma cell lines showed extremely strong bands in Northern blot and RT-PCR/Southern analysis. The enzyme activity was also very high in melanomas as expected. Neuroblastoma and astrocytoma lines showed relatively low levels of the gene expression, whereas they expressed high levels of GD2. Although the mRNA level of the GD3 synthase gene and enzyme activity in individual cell lines correlated positively, some cell lines showed much higher activity than expected from the mRNA level. Among leukaemia lines, adult T cell leukaemia-associated (HTLV-I+) lines showed fairly high levels of the mRNA. On the other hand, T-ALL lines showed very low levels. In addition, GD3 and GD2 expression and mRNA level of the gene during T lymphocyte activation were analysed. Only GD3 expression was induced by any of the stimulatory reagents used, and corresponding up-regulation of the GD3 synthase gene was shown in RT-PCR/Southern analysis.

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Authors and Affiliations

  1. Department of Oncology, Nagasaki University School of Medicine, 1-12-4 Sakamoto, 852, Nagasaki, Japan

    Shuji Yamashiro, Masahiko Okada, Masashi Haraguchi, Keiko Furukawa, Hiroshi Shiku & Koichi Furukawa

  2. Immunology Program, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, 10021, New York, USA

    Kenneth O. Lloyd

  3. Department of Internal Medicine II, Mie University School of Medicine, Edobashi, 514, Tsu, Mie, Japan

    Hiroshi Shiku

Authors
  1. Shuji Yamashiro
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  2. Masahiko Okada
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  3. Masashi Haraguchi
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  4. Keiko Furukawa
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  5. Kenneth O. Lloyd
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  6. Hiroshi Shiku
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  7. Koichi Furukawa
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Additional information

Abbreviations: α2,8S-T, α2,8sialyltransferase; RT-PCR, reverse transcription polymerase chain reaction; HTLV-I, human T lymphotropic virus type I; T-ALL, T cell acute lymphoblastic leukaemia; β1,4GalNAc-T, β1,4-N-acetylgalactosaminyltransferase; PMSF, phenylmethylsulfonylfluoride; NeuAc,N-acetylneuraminic acid; FBS, fetal bovine serum; DMEM, Dulbecco's modified Eagle's medium; mAb, monoclonal antibody; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; Con A, concanavalin A, PHA; phytohaemagglutinin; TPA, 12-O-tetradecanoylphorbol-13-acetate; ATL, adult T cell leukaemia. Ganglioside nomenclature is based on that of Svennerholm [26].

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Yamashiro, S., Okada, M., Haraguchi, M. et al. Expression of α2,8-sialyltransferase (GD3 synthase) gene in human cancer cell lines: high level expression in melanomas and up-regulation in activated T lymphocytes. Glycoconjugate J 12, 894–900 (1995). https://doi.org/10.1007/BF00731251

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  • DOI: https://doi.org/10.1007/BF00731251

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