Ovarian cancer is the leading gynaecological cause of death in women. Altered glycosylation is a feature of ovarian cancer tumour progression and metastasis with increased levels of high mannose structures and bisecting N-glycans found in cancerous cells when compared to non-cancerous cells. The enzyme that is responsible for forming bisecting N-glycans is β 1,4-N-acetylglucosaminyltransferase III (MGAT3) which, due to the nature of the glycosylation pathway, prevents further branching of glycan structures.
To date the majority of cell-based assays use cells grown in the traditional two-dimensional (2D) monolayer cultured on a flat and rigid surface. The move to three-dimensional (3D) cell culture has become increasingly popular in the research community as it allows studies of cellular responses in settings that more closely resemble the in vivo environment. Growing cells on biological scaffolds, eg. decellularised whole organs, in which the extracellular matrix (ECM) has been left intact, provides a 3D cell growth. ECM also contains hormones and growth factors which are important to a cell’s ongoing growth and survival.
In this project we investigated the changes in cell physiology and glycan expression on cell membrane proteins of cells grown in three different methods: traditional 2D flat cell culture, 3D spheroid cell culture and 3D tissue scaffold culture. We analysed differences in expressed glycosylation on proteins from three ovarian cancer cell lines and their MGAT3 deleted counterparts. The differences in specific glycan expression between the different cell culture systems were imaged by fluorescent lectins (eg. E-PHA lectin specific for bisecting GlcNAc).