ganglioside--gd1a and Neoplasm-Metastasis

Mouse FBJ virus-induced osteosarcoma FBJ-S1 cells rich in GD1a are not readily metastatic, whereas FBJ-LL cells with low levels of GD1a are highly metastatic. GD1a was previously shown to suppress metastasis of mouse FBJ cells and to upregulate caveolin-1 and stromal interaction molecule 1 expression. The present study demonstrates that matrix metalloproteinase-9 (MMP-9) expression renders FBJ-LL cells invasive. MMP-9 is inversely regulated by GD1a, based upon four observations: MMP-9 mRNA content was 5 times higher in FBJ-LL cells than FBJ-S1 cells; a GD1a-re-expressing FBJ-LL cell variant produced through beta1,4GalNAcT-1 cDNA transfection expressed lower levels of MMP-9; exogenous addition of GD1a to FBJ-LL cells decreased MMP-9 production in a dose- and time-dependent manner; and treatment of GD1a-rich cells with D-PDMP or siRNA targeting St3gal2 decreased GD1a expression, but augmented MMP-9 expression. This is the first report demonstrating that GD1a negatively regulates expression of MMP-9 at the transcriptional level.

Topics: Animals; Cell Line, Tumor; Cell Movement; Dose-Response Relationship, Drug; Gangliosides; Gene Expression Regulation; Matrix Metalloproteinase 9; Mice; Morpholines; Neoplasm Invasiveness; Neoplasm Metastasis; Osteosarcoma; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; RNA, Small Interfering; Transcription, Genetic

Ganglioside GD1a, which is highly expressed in poorly metastatic FBJ-S1 cells, has been shown to inhibit the serum-induced migration capability of highly metastatic FBJ-LL cells. In the present study, the capacity of FBJ-S1 cells to adhere to vitronectin was found to be about half that of FBJ-LL cells. Pre-treatment of FBJ-LL cells with GD1a decreased this capacity by 30% that of the control, whereas GM1-pre-treatment caused only a 10% decrease, indicating that GD1a specifically inhibits FBJ-LL cell adhesion to vitronectin. Since FBJ-LL cells contain almost no GD1a, transfectants capable of expressing GD1a to varying degrees were produced in this study by transfection of FBJ-LL cells with GM2/GD2-synthase cDNA. Decrease in the serum-induced migration capacity of these transfectants was accompanied by an increment in GD1a expression. Adhesion of the transfectants to vitronectin decreased by 30% as compared with mock-transfected cells. Within 4 to 5 weeks after GD1a-expressing transfectant and mock-transfected cells were transplanted into mice, metastatic nodules were observed in liver, lung, kidney and adrenal glands of mock-transplanted mice, but not in those with GD1a-expressing transfectants, indicating that GD1a suppresses the metastasis of FBJ-osteosarcoma cells, possibly by inhibiting cell migration and cell adhesion. The involvement of the ganglioside in the suppression of metastasis is clearly demonstrated in the present study.

Topics: Animals; Cell Adhesion; Cell Movement; DNA, Complementary; Fibronectins; G(M1) Ganglioside; G(M3) Ganglioside; Gangliosides; Laminin; Mice; Mice, Inbred BALB C; N-Acetylgalactosaminyltransferases; Neoplasm Metastasis; Polypeptide N-acetylgalactosaminyltransferase; Transfection; Tumor Cells, Cultured; Vitronectin

For clarification of the functions of gangliosides on tumor metastasis, examination was made of the ganglioside patterns of poorly metastatic FBJ-S1 and highly metastatic FBJ-LL cells. FBJ-S1 cells expressed GM3 and GD1a, whereas FBJ-LL cells expressed GM3 and slightly expressed GD1a. The capacity for FBJ-LL cells to migrate was ten times that of FBJ-S1 cells, but decreased by a half by pretreatment with GD1a. GD1b or GT1b had the same effect as GD1a, and synthetic sialyl compounds to a lesser extent, suggesting that gangliosides contain more than two sialyl residues to inhibit the migration of FBJ-LL cells.

Topics: Animals; Chromatography, Thin Layer; Gangliosides; Mice; Mice, Inbred BALB C; Neoplasm Metastasis; Tumor Cells, Cultured

The aim of the present study was to investigate the ganglioside expression of the highly metastatic murine lymphoreticular tumour cell line MDAY-D2. Cells were propagated under controlled pH conditions and oxygen supply in bioreactors of 1 and 7.5 l volumes by repeated batch fermentation. Gangliosides were isolated from 2.7 x 10(11) cells, purified by silica gel chromatography and separated into mono- and disialoganglioside fractions by preparative DEAE anion exchange high performance liquid chromatography. Individual gangliosides were obtained by preparative thin layer chromatography. Their structural features were established by immunostaining, fast atom bombardment and gas chromatography mass spectrometry. In addition to gangliosides of the GM1a-pathway (GM2, GM1a and GD1a) and GM1b (IV3Neu5Ac-GgOse4Cer) and GalNAc-GM1b of the Gm1b-pathway, the disialoganglioside GD1 alpha (IV3Neu5Ac, III6Neu5Ac-GgOse4Cer) was found in equal amounts compared to GD1a (IV3Neu5Ac, II3Neu5Ac-GgOse4Cer). All gangliosides were substituted with C24:0, 24:1 and C16:0 fatty acids, sphingosine and N-acetylneuraminic acid as the sole sialic acid.

Topics: Animals; Carbohydrate Sequence; Cell Line; Chromatography, High Pressure Liquid; Chromatography, Ion Exchange; Chromatography, Thin Layer; Gangliosides; Gas Chromatography-Mass Spectrometry; Lymphoma; Mice; Molecular Sequence Data; Neoplasm Metastasis; Neuraminidase; Oligosaccharides; Sialic Acids; Spectrometry, Mass, Fast Atom Bombardment; Tumor Cells, Cultured

Glycolipids of murine lymphoma cell lines with low metastatic (Eb) and high metastatic (ESb) potentials have been investigated. The Eb cell line was characterized by a high quantity of gangliotriaosylceramide (Gg3), gangliotetraosylceramide (Gg4), GM1b, and a new type of disialoganglioside, termed GD1 alpha. In contrast, the high metastatic ESb cell line was characterized by the absence of these glycolipids and instead by the presence of GM3, GM2, GM1a, GD1a, and GD1b gangliosides. A clear cell surface reactivity with monoclonal antibody anti-Gg3 (2D4) was observed only in Eb cells. Thus, Eb cells are distinct from ESb cells in their ability to add the GalNAc residue to LacCer, supplying Gg3 for synthesis of a series of glycolipids via an asialogangliotetraosyl pathway, while ESb cells are capable of synthesizing GM3, which initiates synthesis of ganglio-series gangliosides GM2, GM1a, GD1a, and GD1b. While disialogangliosides of ESb cells were identified as GD1a and GD1b, a disialoganglioside isolated from Eb cells was characterized as having a novel structure (referred to as GD1 alpha) as follows: (formula; see text) Thus, Eb and ESb cells are clearly different in their qualitative sialylation patterns, i.e., the position of sialic acid residues. Cell surface labeling with galactose-oxidase/NaB[3H]4 revealed a high exposure of Gg3 and Gg4 at the Eb cell surface, while both labels were absent in ESb cells. In contrast, ESb cells showed a substantial label at GM1a, which was greatly enhanced after sialidase treatment.

Topics: Animals; Antibodies, Monoclonal; Carbohydrate Sequence; Chromatography, Thin Layer; Gangliosides; Glycolipids; Lymphoma; Membrane Lipids; Mice; Neoplasm Metastasis; T-Lymphocytes