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Glycosphingolipids of Lectin-resistant Mutants of the Highly Metastatic Mouse Tumor Cell Line, MDAY-D2¹

Division of Cancer Research, Mount Sinai Hospital Research Institute, Toronto, Ontario M5G 1X5 [S. F., J. W. D.]; the Cancer Research Center, La Jolla Cancer Research Foundation, La Jolla, California 92037 [M. N. F., M. F.]; and the Department of Biochemistry, Imperial College of Science and Technology, London, England [A. D.]

Neutral and acidic glycolipids in MDAY-D2, a highly metastatic murine tumor cell line, were examined and compared with glycolipids of MDW4 and D33W25-1, two lectin-resistant mutants of MDAY-D2 from distinct genetic complementation classes. D33W25-1 remained highly metastatic while MDW4 cells were found to be nonmetastatic (Dennis, J. W., Donaghue, T., Florian, M., and Kerbel, R. S., Nature (Lond.), 292: 242–245, 1981 and Dennis, J. W. et al., Cancer Res., 46: 4594–4600, 1986). Glycolipid structures were identified by fast-atom bombardment mass spectrometry, methylation analysis, exoglycosidase treatment, and immunostaining. The metastatic MDAY-D2 was found to contain G_M3, G_M2, IV³GalNAc-G_M1b, and high levels of G_M1a, G_M1b, and G_D1a. MDW4 showed a 3-fold decrease in total ganglioside content compared to MDAY-D2 and a corresponding increase in the precursor, glucosylceramide. MDW4 was deficient in G_M1 and accumulated G_M2 and NeuNG-G_M2, indicating a lack of gangliosides having NeuNAc2-3Galß1-3 terminal sequence. Neosynthesis of G_D3 was also observed in MDW4. The metastatic mutant D33W25-1 had a similar pattern of gangliosides as that found in MDAY-D2 cells with N-glycolyl rather than N-acetyl neuraminic acid. These results suggest that (a) the metastatic property of these cell lines may be related to the level of ganglioside, and that (b) the substitution of N-glycolyl for N-acetyl neuraminic acid does not reduce metastatic capacity.

¹ This work was supported by grants from the National Cancer Institute of Canada and the Medical Research Council of Canada (J. W. D.), the National Cancer Institute grant CA-28896 (M. F.), and the Medical Research Council of the United Kingdom (A. D.).

² Recipient of a postdoctoral fellowship from the National Cancer Institute of Canada.

³ Research Scholar of the National Cancer Institute of Canada. To whom requests for reprints should be addressed, at Division of Cancer Research, Mount Sinai Hospital Research Institute, 600 University Avenue, Toronto, Ontario, Canada M5G 1X5.

Received 5/ 2/86. Revised 8/ 4/86. Accepted 9/17/86.

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