Microscale analysis of glycosphingolipids by TLC blotting/secondary ion mass spectrometry: A novel blood group A-active glycosphingolipid and changes in glycosphingolipid expression in rat mammary tumour cells with different metastatic potentials

The glycosphingolipid compositions of rat mammary tumour cell lines with different metastatic potentials for the lung [a parental tumour cell line (MTC) and its subclones MTLn2 (a non metastatic subclone) and MTLn3 (a subclone with high metastatic potential to the lung)] were studied using a newly developed TLC blotting/secondary ion mass spectrometry system and crude glycosphingolipids obtained from 0.5-1 x 10(7) cells of each cell line. G(M3) and G(M2) were the major components of the MTC cell line, but they were very minor components in the MTLn2 and MTLn3 cell lines, G(D1a) being the major ganglioside. HexNAc-fucosyl-G(M1a) was found in the MTLn2 cells by the TLC blotting/SIMS method and the terminal sugar linkage was shown to be a blood group A-type structure by immunostaining. These findings suggest that the ganglioside is a novel type of blood group A-active ganglioside, GalNAc alpha 1-3(Fuc alpha 1-2)G(M1a). No blood group A-active lipid was present in MTLn3 cells, whereas Hex-G(M1a) and neutral glycosphingolipids with more than 5 sugar residues were.