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J. Biochem, 1993, Vol. 114, No. 2 186-193
© 1993 Japanese Biochemical Society

other

Characterization of Ganglioside Expression in Human Melanoma Cells: Immunological and Biochemical Analysis1

Ikuo Kawashima*, Hideki Ozawa*, Masaharu Kotani*, Minoru Suzuki**, Takehiro Kawan**, Makoto Gomibuchi*** and Tadashi Tai*

*Department of Tumor Immunology Honkomagome, Bunkyo-ku, Tokyo 113
**Department of Membrane Biochemistry, Tokyo Metropolitan Institute of Medical Science Honkomagome, Bunkyo-ku, Tokyo 113;
***Department of Thoracic Surgery, Nippon Medical College Sendagi, Bunkyo-ku, Tokyo 113

The expression of N-glycolylneuraminic acid (NeuGc)-containing gangliosides in human melanoma cells grown both in culture and as xenografts in athymic (nu/nu) mice was analyzed extensively with specific mouse monoclonal antibodies (MAbs). Three MAbs (GMR8, GMR14, and GMR3) specific for GM3(NeuGc), GM2(NeuGc), and GD3(NeuGc-NeuGc-), respectively, were used. Significant differences were observed in the ganglioside compositions between the cultured cells in vitro and the tumors grown in vivo The major difference was that the cells cultured in serum-free medium did not express any NeuGc-containing gangliosides, whereas those grown in nude mice expressed a number of NeuGc-containing gangliosides, namely GM3(NeuGc), GM2(NeuGc), GD3(NeuAc-NeuGc-), GD3(NeuGc-NeuAc-), and GD3(NeuGc-NeuGc-). The structures of these gangliosides were also determined chemically. No activity of CMP-NeuAc hydroxylase was demonstrated either in the melanoma cells cultured in vitro or in those grown in nude mice, suggesting that these cells incorporated NeuGc-containing glycoconjugates from the mouse sera and converted them to other NeuGc-containing gangliosides. The mouse sera contained only GM2(NeuGc), but not the other NeuGc-containing gangliosides or any NeuAc-containing gangliosides.

1This work was supported in part by a Grant-in-Aid for Scientific Research on Priority Areas (004250101) from the Ministry of Education, Science and Culture of Japan and by a grant from the Naito Foundation.


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