Investigations into altered cellular glycosylation associated with metastatic competence in breast cancer cell lines
type: CreativeWork
description: Metastasis is the main cause of cancer patient death owing to the fact that metastatic disease is difficult to treat. Aberrant glycosylation is characteristic of cancer and plays a role in metastatic mechanisms. HPA, a lectin from the Roman snail Helix pomatia, recognises glycans terminating in α-GalNAc, and is associated with aggressive biological behaviour, poor patient prognosis and metastasis in breast and other cancers. Three breast cancer cell lines characterised to stably synthesise an array of GalNAc-glycans were employed in the work reported here (MCF7, ZR751 and BT474). HPA lectin labelling and quantification were optimised and it revealed a more complex HPA-binding profile than previously reported. It was observed that HPA-positive cells were rounded, reminiscent of epithelial cells, and HPA-negative cells were elongated, and reminiscent of mesenchymal cells. One of the aberrant glycans that HPA reportedly binds is the initial O-glycan Tn structure which is commonly exposed in cancer. qPCR was performed to assess the gene expression levels of O-linked glycosylation initiation enzymes, GALNTs as well as C1GalT and its molecular chaperone COSMC. This analysis revealed that there is no clear association between aberrant glycosylation recognised by HPA-binding, although down-regulation of C1GalT and COSMC may result in failed normal chain extension. A method of isolating cells based on their HPA-binding profile was developed. Cells were separated and grown on post-separation for 3 days. It was observed that the glycosylation profiles of the HPA-positive cell population and the HPA-negative cell population reverted to a mixed population over time that was similar to the unseparated lectin labelling proportions. The cell morphologies also reverted. The plasticity observed was reminiscent of EMT. Microarray analysis revealed that several EMT-associated genes were differentially expressed between the HPA-positive and HPA-negative isolated cell populations. Using …
abstract: Metastasis is the main cause of cancer patient death owing to the fact that metastatic disease is difficult to treat. Aberrant glycosylation is characteristic of cancer and plays a role in metastatic mechanisms. HPA, a lectin from the Roman snail Helix pomatia, recognises glycans terminating in α-GalNAc, and is associated with aggressive biological behaviour, poor patient prognosis and metastasis in breast and other cancers. Three breast cancer cell lines characterised to stably synthesise an array of GalNAc-glycans were employed in the work reported here (MCF7, ZR751 and BT474). HPA lectin labelling and quantification were optimised and it revealed a more complex HPA-binding profile than previously reported. It was observed that HPA-positive cells were rounded, reminiscent of epithelial cells, and HPA-negative cells were elongated, and reminiscent of mesenchymal cells. One of the aberrant glycans that HPA reportedly binds is the initial O-glycan Tn structure which is commonly exposed in cancer. qPCR was performed to assess the gene expression levels of O-linked glycosylation initiation enzymes, GALNTs as well as C1GalT and its molecular chaperone COSMC. This analysis revealed that there is no clear association between aberrant glycosylation recognised by HPA-binding, although down-regulation of C1GalT and COSMC may result in failed normal chain extension. A method of isolating cells based on their HPA-binding profile was developed. Cells were separated and grown on post-separation for 3 days. It was observed that the glycosylation profiles of the HPA-positive cell population and the HPA-negative cell population reverted to a mixed population over time that was similar to the unseparated lectin labelling proportions. The cell morphologies also reverted. The plasticity observed was reminiscent of EMT. Microarray analysis revealed that several EMT-associated genes were differentially expressed between the HPA-positive and HPA-negative isolated cell populations. Using …
author: Beaman, Ellie-May
language: en
date published: 2017
date modified: 2026-07-24T03:43:15Z
provider: Oxford Brookes University
publisher: Networked Digital Library of Theses and Dissertations
type: WebPage
source organization: Oxford Brookes University

PropertyValue
property ID: DOI
value: 10.24384/p563-y105