Glycoconjugates of murine tumor lines with different metastatic capacities. I. Differences in fucose utilization and in glycoprotein patterns

Abstract

Since various experimental findings point towards an important role of cell surface carbohydrates - in particular sialic acid - in cancer metastasis, the rationale of this study was to look for possible differences in carbohydrate metabolism and glycoprotein expression in well-defined related tumor lines of different metastatic capacity. The tumor lines analyzed were L5178Y E (= Eb), a low-metastasizing, methylcholanthrene-induced lymphoma of a DBA/2 mouse, and L5178Y ES (= ESb), a spontaneous high-metastatic variant thereof. A non-related, highly metastasizing tumor, MDAY-D2, and ConA-stimulated spleen cells were included in the study. These cell lines were compared for incorporation rates of various labelled carbohydrates and for glycoprotein patterns in SDS-polyacrylamide gels. Marked differences were observed in the incorporation of 3H-fucose, while the incorporation of 3H-galactose and 3H-mannose was similar in the different cell lines studied. Only the metastatic variant ESb incorporated 3H-fucose at a rate similar to that of ConA-stimulated T-cell blasts. Eb cells did not incorporate 3H-fucose while MDAY-D2 had a significantly lower 3H-fucose incorporation rate. Separation and purification of the intracellular products of 3H-fucose by gel filtration and high-voltage electrophoresis revealed in Eb cells a block in the synthesis of fucosylated glycoproteins at the step of the fucose-I-P-guanylyltransferase. No apparent defect in the fucose pathway was detectable in MDAY-D2 cells. An Eb----ESb shifted cell line regained the ability to incorporate 3H-fucose. All tumors displayed unique glycoprotein patterns in SDS-PAGE. Labelling with 3H-mannose revealed the most distinct bands, while labelling with 3H-galactose gave fewer and broader bands. Although clonal instability of metastasizing tumor variants has been frequently reported, subclones of Eb and ESb showed characteristics similar to those of the original cell lines with regard to metastatic capacity, fucose metabolism and glycoprotein expression. These results will be discussed in relation to differences in fucose metabolism and in surface expression of fucose as observed in other tumor systems consisting of high- and low-metastatic lines.

Expression and enhanced secretion of proteochondroitin sulphate in a metastatic variant of a mouse lymphoma cell line

“…These tumour lines have been extensively investigated concerning their invasive capacity (Schirrmacher et al, 1979b), their immunogenicity and tumour-associated transplantation antigens (Schirrmacher et al, 1979a) and their differentiation antigens (Altevogt et al, 1982). The glycoproteins and glycolipids of cell lines Eb and ESb have been described in two preceding studies (Schwartz et al, 1984;Schwartz et al, 1985). In the present work, we demonstrate that GAGs from highly metastatic ESb cells are essentially different compared to those from low metastatic Eb cells.…”

“…Eb and ESb-cells are distinguished from each other by their specific expression of differentiation antigens (Altevogt et al, 1982), tumour-associated transplantation antigens (TATA) (Schirrmacher et al, 1979a) and cell surface glycoconjugates (Schwartz et al, 1984 whereas the variant ESb-MP is more closely related to ESb. Both cell types express identical differentiation antigens and TATA (Fogel et al, 1983).…”

Determination of cell-free interleukin 2 receptor level in the serum of normal animals and of animals bearing IL-2 receptor positive tumours with high or low metastatic capacity

“…The spontaneous high metastatic variant ESb is most likely derived from a spontaneous fusion of the parental Tlymphoma Eb with a host macrophage (Larizza et al, 1984). The two lines were found to differ in many cell surface properties including expression of differentiation antigens , of composition of membrane glycoproteins (Schwartz et al, 1984), glycolipids (Murayama et al, 1986) and in the shedding of membrane vesicles (Barz et al, 1985;Schirrmacher & Barz, 1986 (Waller et al, 1986). In vivo, however, the ESb-M line showed greatly decreased overall malignancy (Benke et al, 1987).…”

Glycosphingolipid expression on murine L1-fibrosarcoma cells: analysis of clonal in vivo and in vitro selected sublines with different lung colonisation potential

“…Most of the relevant evidence is indirect suggesting that membrane constituents which are less expressed on non-metastatic cells or even absent may be responsible for the properties of metastatic tumour cells. Accordingly, high and low metastatic cells differ in cell surface sialic acid (Yogeeswaran et al, 1981b) in glycolipid (Schwartz et al, 1985;Laferte et al, 1987) and in glycoprotein composition (Schwartz et al, 1984;Steck et al, 1987).…”