The neural cell adhesion molecule, NCAM, plays an important role in cell-cell adhesion. Therefore, we have studied NCAM expression in the glioma cell lines BT4C and BT4Cn. We demonstrate that the 2 cell lines differ in their metastatic ability; while BT4C cells have a very low capacity for producing experimental metastases, that of BT4C, cells is high. In BT4C cells NCAM is synthesized as 4 polypeptides with M,'s of 190,000, 140,000, I15,000 and 97.000. The 140,000, I15,OOO and 97,000 polypeptides are glycosylated and for the 140,000 and I 15,000 polypeptides sulfatation is observed. Conversely, no NCAM protein synthesis is observed in BT4C, cells, even though NCAM mRNA is expressed, Thus, development of an increased metastatic capacity is accompanied by the disappearance of NCAM protein expression in this model system. The functional importance of NCAM expression was studied by a cell-substratum binding assay in which the binding of BT4C and BT4C, cells to NCAM immobilized to glass was assessed. We found that BT4C cells adhere specifically to NCAM, and that adhesion is inhibited by anti-NCAM Fab'-fragments, while no specific binding of BT4Cn cells to NCAM was observed. The BT4C and BT4Cn cell lines thus constitute an important new model system for the study of tumor invasion and metastasis and of the role of cell adhesion molecules in these processes.
MATERIAL AND METHODS
Cell cultures
Cells were cultured in 10% fetal calf serum (Gibco, Paisley, UK) in Dulbecco's modified Eagle's medium (DME; Gibco) at 37ยฐC in a humidified atmosphere with 5% CO,.