Control of metastasis by Asn-linked, beta1-6 branched oligosaccharides in mouse mammary cancer cells.

Studies in cell lines and malignant human tissues have shown that increased cell-surface Asn-linked beta1-6(GlcNAcbeta1-6Man) branching is associated with increased tumorigenic and metastatic properties. In this study, three mouse mammary cancer cell lines were transfected with an expression vector containing the mouse cDNA for N-acetylglucosaminyltransferase V (GlcNAcT-V EC 2.4.1.155), the glycosyltransferase responsible for initiating beta1-6 branching on Asn-linked carbohydrates. The cell lines were screened for increased cytotoxicity to L-PHA, a lectin specific for beta1-6 branching structures. Cell lines exhibiting increased L-PHA cytotoxicity expressed increased levels of beta1-6 branching structures. Northern blots detected the presence of GlcNAcT-V transcribed from the expression vector in the L-PHA sensitive cell lines. After injection into the tail veins of mice, transfected cell lines with increased beta1-6 branching on the cell surface formed elevated levels of lung tumors relative to control transfected cell lines (P < 0.002). Western blots of membrane proteins from GlcNAcT-V transfected and control cells probed with the lectins DSA and WGA did not show an increase in polyN-acetyllactosamine and sialic acid content in the transfected cell lines. These results demonstrate that a specific increase in beta1-6 branching due to an elevation in GlcNAcT-V expression increases metastatic potential.

N-linked oligosaccharides and metastatic propensity in in vivo selected mouse mammary adenocarcinoma cells.

Studies using metastatic variant selected in vivo from a cloned parental cell line demonstrate that the expression of beta1-6 branched, N-linked carbohydrates and sialic acid were positively associated with in vitro invasiveness and inversely associated with metastatic potential, adherence, and in vivo growth rate. These results suggest that at least within one tumor model, a negative association occurs between metastatic potential and 1-6 branched oligosaccharide expression. In these studies two metastatic variants, Cl-66M1 and Cl-66M2, were selected following serial in vivo passage of Cl-66, a clonal cell line obtained from a mouse mammary adenocarcinoma cell line. The parent cell line and the two metastatic variants were approximately equal in their adherence to fibronectin, laminin, and collagen type IV coated plastic. In contrast, both Cl-66M1 and Cl-66M2 had a significantly increased ability to invade through matrigel invasion chambers and expressed significantly increased levels of beta1-6 branched, N-linked carbohydrates, and sialic acid compared to the clonal parental cell line, Cl-66. Furthermore, the in vivo tumor growth rates of these selected variants were decreased compared to Cl-66 with the longest tumor volume doubling time observed with Cl-66M2.