Syndecan-1 regulates FGF8b responses in S115 mammary carcinoma cells.

In murine mammary carcinoma cells Shionogi 115 (S115) testosterone induces phenotypical transformation which is largely due to expression of fibroblast growth factor (FGF) 8b. Concomitantly, the expression of the cell surface heparan sulfate proteoglycan syndecan-1 is down-regulated. However, if syndecan-1 expression is maintained by transfection with a testosterone-driven syndecan-1 construct, transformation does not occur. Here we have investigated how the down-regulation of syndecan-1 expression in testosterone-treated S115 cells and the high level of expression in syndecan-1 transfected cells influence the cellular responses toward FGF8b. Our results show that high level of syndecan-1 is associated with a decreased magnitude and duration of the FGF8b induced Erk phosphorylation. This effect was observed regardless whether the cells were stimulated directly with exogenous FGF8b or with testosterone to induce autocrine FGF8b production. Moreover, syndecan-1 transfected cells did not respond to FGF8b stimulation by increase in the intracellular free calcium, whereas untransfected cells displayed a rapid (10 s) induction. These data suggest that, in S115 cells, syndecan-1 acts as a modulator of FGF8b signaling that can limit cellular responses to FGF receptor activation. The decreased levels of syndecan-1 expression and upregulation of the FGF signaling system seen in many cancers may contribute to the proliferation of the malignant cells in vivo.

Expression and characterization of minican, a recombinant syndecan-1 with extensively truncated core protein.

Syndecan-1 is an integral membrane heparan sulfate/chondroitin sulfate proteoglycan, involved in the control of cell growth and differentiation. The biological activities of syndecan-1 involve interactions with a variety of extracellular ligands, such as growth factors and matrix components, that are mainly mediated by the heparan sulfate moieties. The expression of syndecan-1 is downregulated in various malignant tumors, and low levels of expression appear to correlate with poor prognosis of some cancer types. On the other hand, the extracellular portion of syndecan-1 (ectodomain) has been demonstrated to inhibit the proliferation of various cancer cells in culture, suggesting that proteoglycan-like molecules should be studied further with regard to their antitumor activities. We have expressed, in CHO cells, a truncated syndecan-1 ectodomain ("minican") harboring domains for glycosaminoglycan attachment and antibody recognition. Analysis of recombinant minican indicates that it shares some of the biochemical and biological characteristics attributed to syndecan-1 ectodomain. Minican was thus substituted with heparan sulfate chains and bound to extracellular matrix proteins as well as fibroblast growth factors. Notably, minican inhibited the proliferation of S115 mouse mammary carcinoma cells and the effect seemed to involve inhibition of the Ras/Erk signaling pathway. Our data suggest that recombinant syndecan-1 with a minimal protein component is biologically active. This information may provide useful in further design of proteoglycan-like antitumor molecules.