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Title: Developing and using atomic force microscopy to investigate mechanisms of tumour metastasis
Author: Albon, Jacob George
ISNI:       0000 0004 5350 6566
Awarding Body: University of Sheffield
Current Institution: University of Sheffield
Date of Award: 2015
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Mortality in breast cancer is increasingly linked to metastasis to bone, which is currently incurable and the mechanisms involved in this process are not fully understood. By using atomic force microscopy to extract the mechanical and adhesive properties of metastatic breast cancer cells under near physiological conditions the aim of the work presented in this thesis is to develop the methods used to extract these properties as well as enhance understanding of tumour metastasis. The biomechanical properties of three human breast cell lines with varying metastatic potential have been investigated through atomic force microscopy based indentation, using three separate analytical models. The indentation studies were used to reveal mechanical properties of living cells which can be attributed to cellular structures such as the cytoskeleton, cell membrane, cytoplasm and numerous intracellular organelles. This is the first study to compare three contact mechanical models on these cells. The results demonstrate that benign MCF-10A breast cells are less deformable than the invasive MCF-7 and metastatic MDA-MB-231 breast cancer cell lines. Force maps have also revealed elastic heterogeneity across the cell surface of all three cell lines which appears to correlate with intracellular structures. The results demonstrate that whole cell mapping has the potential to investigate cell mechanics using a variety of approaches. The biomechanical properties of sub-populations of the MDA-MB-231 cell line were also investigated. Sub-populations of cancer cells which were shown to display 'stem-cell-like' properties and 'normal' cancer cells which did not were identified using a number of in vitro techniques and selected for AFM indentation experiments via fluorescence microscopy. The results indicated no significant difference between the measured mechanical properties of the 'stem-like' and 'normal' cancer cells. Adhesive interactions between metastatic MDA-MB-231 cells and 'osteoblast-like' Saos-2 cells were then characterised using an AFM based single cell force spectroscopy (SCFS) protocol. The detachment force required to separate the two cell types was measured for untreated, zoledronic acid treated and anti-N-cadherin treated cells. The results showed that both treatments significantly reduced the detachment force. This result is important in understanding the mechanisms by which adjuvant zoledronic acid therapy may reduce bone resorption and metastases in patients with advanced breast cancer.
Supervisor: Hobbs, Jamie K. ; Brown, Nicola J. Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available