MUC1 O-glycosylation contributes to anoikis resistance in epithelial cancer cells.

Anoikis is a fundamental cellular process for maintaining tissue homeostasis. Resistance to anoikis is a hallmark of oncogenic epithelial-mesenchymal transition and is a pre-requisite for metastasis. Previous studies have revealed that the heavily glycosylated mucin protein MUC1, which is overexpressed in all types of epithelial cancer cells, prevents anoikis initiation in response to loss of adhesion. This effect of MUC1 is largely attributed to its extracellular domain that provides cell surface anoikis-initiating molecules with a 'homing' microenvironment. The present study investigated the influence of O-glycosylation on MUC1 extracellular domain on MUC1-mediated cell resistance to anoikis. It shows that stable suppression of the Core 1Gal-transferase (C1GT) by shRNA substantially reduces O-glycosylation in MUC1-positively transfected human colon cancer HCT116 cells and in high MUC1-expressing SW620 cells. Suppression of C1GT significantly increased anoikis of the MUC1-positive, but not MUC1-negative, cells in response to suspended culture. This effect was shown to be associated with increased ligand accessibility to cell surface anoikis-initiating molecules such as E-cadherin, integrinß1 and Fas. These results indicate that the extensive O-glycosylation on MUC1 extracellular domain contributes to MUC1-mediated cell resistance to anoikis by facilitating MUC1-mediated prohibition of activation of the cell surface anoikis-initiating molecules in response to loss of cell adhesion. This provides insight into the molecular mechanism of anoikis regulation and highlights the importance of cellular glycosylation in cancer progression and metastasis.

Interaction of galectin-3 with MUC1 on cell surface promotes EGFR dimerization and activation in human epithelial cancer cells.

Epidermal growth factor receptor (EGFR) is an important regulator of epithelial cell growth and survival in normal and cancerous tissues and is a principal therapeutic target for cancer treatment. EGFR is associated in epithelial cells with the heavily glycosylated transmembrane mucin protein MUC1, a natural ligand of galectin-3 that is overexpressed in cancer. This study reveals that the expression of cell surface MUC1 is a critical enhancer of EGF-induced EGFR activation in human breast and colon cancer cells. Both the MUC1 extracellular and intracellular domains are involved in EGFR activation but the predominant influence comes from its extracellular domain. Binding of galectin-3 to the MUC1 extracellular domain induces MUC1 cell surface polarization and increases MUC1-EGFR association. This leads to a rapid increase of EGFR homo-/hetero-dimerization and subsequently increased, and also prolonged, EGFR activation and signalling. This effect requires both the galectin-3 C-terminal carbohydrate recognition domain and its N-terminal ligand multi-merization domain. Thus, interaction of galectin-3 with MUC1 on cell surface promotes EGFR dimerization and activation in epithelial cancer cells. As MUC1 and galectin-3 are both commonly overexpressed in most types of epithelial cancers, their interaction and impact on EGFR activation likely makes important contribution to EGFR-associated tumorigenesis and cancer progression and may also influence the effectiveness of EGFR-targeted cancer therapy.