Had-1, a uridine 5'-diphosphogalactose transport-defective mutant of mouse mammary tumor cell FM3A: composition of glycolipids, cell growth inhibition by lactosylceramide, and loss of tumorigenicity.
Glycolipid compositions of mouse mammary tumor cell FM3A and its Newcastle disease virus-resistant mutant cell, Had-1, which was also characterized as a defective mutant of UDP-galactose transport to Golgi apparatus, have been studied. The major neutral glycolipid in FM3A was Gal beta 1-4Glc beta 1-1Cer (LacCer) (95%) and the rest was Glc beta 1-1Cer. The concentration of neutral glycolipids in Had-1 was only about one-fifth of that in FM3A. GlcB1-1Cer in Had-1 accounted for 79% of neutral glycolipids and the rest was LacCer, the content of which was decreased to 4% of that in FM3A. Ganglioside patterns of the two cell lines were similar, although gangliosides with N-glycolylneuraminic acid were increased in Had-1 cells compared with that in FM3A cells. The presence of NeuAc alpha 2-3-Gal beta 1-4GlcNAc beta 1-3Gal beta 1-4Glc beta 1-1Cer, NeuAc alpha 2-3Gal beta 1-4GlcNAc beta 1-3Gal beta 1-4GlcNAc beta 1-3Gal beta 1-4Glc beta 1-2Cer, GM3, and GD3 was demonstrated by thin-layer chromatography immunostaining. 125I-Labeled Newcastle disease virus bound only poorly to gangliosides extracted from either FM3A or Had-1 cells on a high performance thin-layer chromatography plate. The effects of glycolipids on the growth of the two cell lines were also studied. Had-1 cells were more sensitive to glycolipids added exogenously than FM3A cells. Addition of GM3 had a stimulative effect on cell growth of Had-1. LacCer, Gal beta 1-3GalNAc beta 1-4Gal beta 1-4Glc beta 4-1Cer, and Glc beta 1-1Cer inhibited the growth of Had-1 cells. LacCer was the most potent inhibitor. LacCer immobilized on the culture plate also inhibited the growth of Had-1 cells. The inhibitory effect was recovered completely overcome by transferring the cells to LacCer-free medium. Had-1 cells were not tumorigenic in C3H/He mice, and furthermore the tumorigenic activity of FM3A cells was suppressed by the prior administration of Had-1 cells.
Taki
T
T
Department of Biochemistry, Faculty of Medicine, Tokyo Medical and Dental University, Japan.
Ogura
K
K
Rokukawa
C
C
Hara
T
T
Kawakita
M
M
Endo
T
T
Kobata
A
A
Handa
S
S
D000818:Animals; D015703:Antigens, CD; D002449:Cell Aggregation / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002855:Chromatography, Thin Layer; D005260:Female; D006017:Glycolipids / Q000032:analysis*; D006028:Glycosphingolipids / Q000494:pharmacology*; D007694:Killer Cells, Natural / Q000276:immunology; D007790:Lactosylceramides; D008325:Mammary Neoplasms, Experimental / Q000737:chemistry / Q000473:pathology*; D051379:Mice; D008809:Mice, Inbred C3H; D009154:Mutation; D009368:Neoplasm Transplantation; D009522:Newcastle disease virus / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014531:Uridine Diphosphate Galactose / Q000378:metabolism
D015703:Antigens, CD; D006017:Glycolipids; D006028:Glycosphingolipids; D007790:Lactosylceramides; D014531:Uridine Diphosphate Galactose; C009744:CDw17 antigen