Characterisation of the Helix pomatia agglutinin binding glycoproteins of colorectal cancer cell lines and tissue samples

Saint-Guirons, J. 2007. Characterisation of the Helix pomatia agglutinin binding glycoproteins of colorectal cancer cell lines and tissue samples. PhD thesis University of Westminster School of Life Sciences

TitleCharacterisation of the Helix pomatia agglutinin binding glycoproteins of colorectal cancer cell lines and tissue samples
TypePhD thesis
AuthorsSaint-Guirons, J.
Abstract

Colorectal cancer (CRC) is one of the most common malignancies in the US

and Western Europe and metastatic dissemination after CRC is a leading cause of

mortality. New markers predictive of cancer cell behaviour are actively sought both as targets and as a means of predicting patient prognosis. The lectin from the Roman snail Helix pomatia (HPA) has attracted interest as tool for the detection of metastatic colorectal cancer but the HPA binding partners have remained poorly characterised. We established an in vitro model using human colorectal cancer cell lines

ranging from HPA negative, non metastatic, to HPA positive and metastatic. Confocal microscopy was used to assess the HPA binding pattern in the cell lines and the

monosaccharides N-acetylgalactosamine, N-acetylglucosamine, and sialic acid were used to inhibit the interaction between the lectin and the cancer cells.

A proteomic approach based on cell membrane isolation, pre-fractionation using

lectin affinity chromatography, followed by 2-dimensional electrophoresis and

MALDI-TOF-MS enabled the identification of the HPA binding proteins in the

metastatic cancer cell line HT29. The proteins that eluted in the HPA binding fraction were present either by virtue of their ability to bind directly to HPA or as protein complexes of HPA binding partners and included molecules involved in cell adhesion / migration (integrin a6, integrin aV, annexins) re-modeling (filament proteins including oc tubulin, Ptubulin, cytokeratins, actin) and anti-apoptotic pathways (Hsp-70, Hsp-90, Hsp-96 and TNFR-1). Although many of these proteins have previously been described as altered in cancer, we are not aware of a single reagent like HPA which will

concurrently bind all of these molecules.

Year2007
FileSaint-Guirons.pdf
Publication dates
Completed2007

Related outputs

Proteome analysis of metastatic colorectal cancer cells recognized by the lectin Helix pomatia agglutinin (HPA)
Saint-Guirons, J., Zeqiraj, E., Schumacher, U., Greenwell, P. and Dwek, M. 2007. Proteome analysis of metastatic colorectal cancer cells recognized by the lectin Helix pomatia agglutinin (HPA). Proteomics. 7 (22), pp. 4082-4089.

A proteomic approach to identify the integrins α 6 and α V as HPA binding partners in the metastatic colorectal cancer cell line HT29
Saint-Guirons, J., Zeqiraj, E., Greenwell, P. and Dwek, M. 2006. A proteomic approach to identify the integrins α 6 and α V as HPA binding partners in the metastatic colorectal cancer cell line HT29. Molecular and Cellular Proteomics. 5 (10 (supplement)), p. S148.

Confocal microscopy for the identification of glycosylation changes associated with metastatic colorectal cancer
Zeqiraj, E., Saint-Guirons, J., Kerrigan, M.J.P. and Dwek, M. 2005. Confocal microscopy for the identification of glycosylation changes associated with metastatic colorectal cancer. The Molecular Biology of Colorectal Cancer. UBHT Education Centre, Bristol, UK 10 - 11 Mar 2005

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