E-cadherin is a cell adhesion protein required for epithelial tissue integrity. In many cancer cells mis-regulation of E-cadherin adhesions causes increased progression and invasion of cancer. Alteration in E-cadherin dynamics could therefore serve as an early molecular biomarker of metastasis. In this project, I used E-cadherin FRAP to asses real time dynamics of cadherin junctions in a pancreatic cancer mice model of in a variety of micro-environments. My data showed that p53 mutation drives metastasis through mobilizing E-cadherin in junctions. Also, I used FRAP as a pharmaco-dynamic marker to assess the effect of an anti-invasive drug (dasatinib) in pancreatic tumours in vivo. Moreover, my E-cadherin FRAP data along with cross-linking experiments and disruption of E-cadherin interactions by mutation provided a comprehensive framework for understanding E-cadherin dynamics at cell-cell. Here, I have identified four distinct populations of E-cadherin within regions of cell-cell contact and characterized the interactions governing their mobility using FRAP. These pancreatic cancer cells had the immobile fraction (Fi) of E-cadherin-GFP comprised adhesive and non-adhesive populations. The remaining mobile fraction (Fm) also comprised of non-adhesive and adhesive populations, one population moves at the rate of pure diffusion, and therefore represents free E-cadherin monomers. The other population moves more slowly, and represents E-cadherin monomers turning over within immobile complexes. Inclusion of E-cadherin into either adhesive population requires cis-, trans-, and actin interactions. The signaling pathways in cells dramatically affect the fractions of these cadherin components. I showed that understanding the dynamics of these four populations of E-cadherins could be used to design or interpretation of future pharmacological and genetic experiments to probe the function of E-cadherin in development, disease progression, and response to therapy.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:631001 |
| Date | January 2014 |
| Creators | Rud-Majani, Zahra Erami |
| Publisher | University of Glasgow |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | http://theses.gla.ac.uk/5619/ |
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