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Title: Bioimaging and quantitative analysis of bladder cancer invasion
Author: Koo, V. S. W.
ISNI:       0000 0001 3601 797X
Awarding Body: Queen's University Belfast
Current Institution: Queen's University Belfast
Date of Award: 2008
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TGF-beta1 is known to induce changes tumour morphology and motility which are associated with invasion and metastasis via the phenomenon of epithelial-mesenchymal transition (EMD. Although morphology and motility can be determined in vitro, tumour invasion and metastasis are best investigated in an animal model that can be monitored. The aim of this study was to quantify the effects ofTGF-bT;ta1 on AY-27 bladder tumour and to establish DsRed2 fluorescence monitoring in the AY-27/F344 Fisher rat bladder tumour model using IVIS®200 imaging system. Using cbnventional microscopic assessment, scratch wound and Matrigel assays, we showed that TGF-beta1 induces spindle-shape morphology and significantly increased the motility and invasion in AY-27 cells. Preliminary data showed decreased expression of cytokeratin 18 and polarised distribution of vimentin of TGF-beta1 treated cells, indicating the occurrence of EMT. We also developed the Spindle Index assay that objectively quantified morphology change; and invented a novel chemo-attractant based Koo Assay of Migration which quantified tumour motility. These novel assays concurred with the above reference assays and can serve as an adjunctive investigative tool. DsRed2 was transfected into AY-27 using Lipofectamine2000 and the fluorescent intensity and stability were quantified using IVIS®200 and phase-contrasUfluorescent microscopy. However. the DsRed2 fluorescence intensity was not bright or stable. In addition. the DsRed2 transfection has altered the. morphology and growth characteristics of the cell. Instead, we used tdTomato, a variant of DsRed, and found it superior to DsRed2 in the fluorescent intensity and stability and it did not alter the characteristics of AY-27 cells. We showed that fluorescence detection of SUbcutaneously injected AY-27/tdTomato in F344 rat was successful using IVIS®200. However. intravesical fluorescence detection was hampered due to the thick rat tissue overlying the bladder. We suggest that f1uorescently tagged bladder or other deep intra-abdominal tumour model in rats may not be suitable for monitoring using IVIS®200.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (M.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available