This project was designed to assess clinical and biological parameters of prognosis in high grade breast cancer (BC). The first approach aimed to define Clinicoepidemiological parameters associated with poor survival in a retrospective BC cohort of 1339 non-metastatic patients presenting at Singleton Hospital, South Wales, UK. Median follow-up was 5.4 years (range 0.09-10.14 years). Results of this analysis supported the role of histological grade (HG) as a prognostic factor and a tumour classifier. HG stood as one of the main variables associated with OS, DDFS and DFS with only N3 disease conferring worse prognosis. This analysis contributed with clinical information and survival data required for sample selection for subsequent comparative genomic hybridisation (CGH) studies. Array Comparative genomic hybridisation (aCGH) was performed in 78 cases (67 HG3 and 11 HG2) cases aiming to identify copy number alterations (CNA) associated with poor survival. aCGH protocol optimisation was required to obtain reproducible results and a new simplified aCGH protocol was described. A region of chromosomal gain in Chromosome 5 (5q35.1 to 5q35.2) was significantly associated with Cancer-specific survival (CSS; FDR<0.2). DUSP-1 and MSX2 genes were among those candidate genes validated In-silico for poorer prognosis in BC. FINALLY, THE ISSUE OF TAMOXIFEN RESISTANCE WAS ADDRESSED BY ATTEMPTING TO IDENTIFY CNA IN OESTROGEN RECEPTOR (ER) POSITIVE/TAMOXIFEN TREATED PATIENTS ASSOCIATED WITH EARLY RELAPSE/DEATH ≤5 YEARS (TAMRG) COMPARED WITH A SUB-GROUP OF PATIENTS ALIVE AND WELL AFTER 5 YEARS ON FOLLOW-UP (TAMCG). A REGION OF GAIN ON CHROMOSOME 7 (188219-6234052) WAS ASSOCIATED WITH TMACG (P:0.05) WHICH WAS ALSO ASSOCIATED WITH SIGNIFICANT OVER-EXPRESSION OF SNX8 USING IN-SILICO VALIDATION IN LOI ET AL DATASET. RESULTS FROM THIS STUDY CONTRIBUTE TOWARDS THE IDENTIFICATION OF CANDIDATE PROGNOSTIC GENES IN BC. BIOLOGICAL VALIDATION OF THESE RESULTS IS RECOMMENDED. FURTHER RESEARCH IS NEEDED TO ASSESS THE FUNCTIONAL RELEVANCE OF THESE CANDIDATE GENES.

Background:Chemotherapy resistance is a major obstacle in effective neoadjuvant treatment for locally advanced breast cancer. The ability to predict tumour response would allow chemotherapy administration to be directed towards only those patients who would benefit, thus maximising treatment efficiency. This project aimed to identify predictive protein biomarkers associated with chemotherapy resistance, using proteomic analysis of fresh breast cancer tissue samples. Materials and Methods:Chemotherapy-sensitive (CS) and chemotherapy-resistant (CR) tumour samples were collected from breast cancer patients who received neoadjuvant therapy consisting of epirubicin with cyclophosphamide followed by docetaxel. Comparative proteomic analysis was performed, to identify differentially expressed proteins (DEPs) between CS and CR invasive ductal carcinoma samples, using 2-dimensional polyacrylamide gel electrophoresis (2D-PAGE) with MALDI-TOF/TOF mass spectrometry and antibody microarray analysis. DEPs were submitted to Ingenuity Pathway Analysis (IPA) to identify any canonical pathway links, confirmed using western blotting and clinically validated in a pilot series of archival breast cancer samples, from patients treated with neoadjuvant chemotherapy. Results:Five datasets were generated by antibody microarray analysis, revealing 38 targets. Of these, 7 DEPs were identified in at least 2 datasets and these included 14-3-3 theta/tau, BID and Bcl-xL. Three datasets were generated using 2D-PAGE with MALDI-TOF/TOF MS, containing 132 unique DEPs. These included several isoforms of 14-3-3 proteins. The differential expression of 14-3-3, BID and Bcl-xL was confirmed by immunoblotting in samples used for the discovery phase. Clinical validation using immunohistochemical analysis of archival breast cancers revealed 14-3-3 theta/tau and tBID to be significantly associated with chemotherapy resistance. Discussion:The use of comparative proteomic techniques using fresh clinical tumour samples, for the search for putative biomarkers of chemotherapy resistance has been successful. Two DEPs; 14-3-3 theta/tau and tBID have passed through all stages of the biomarker discovery pipeline, and present themselves as putative predictive biomarkers of neoadjuvant chemotherapy resistance in breast cancer.

In the Western world, breast cancer is the most frequent malignancy in women and still the leading cause of cancer related deaths, therefore, a better understanding of the disease is needed. Adequate therapeutic targets for all breast cancer types have not been identified yet, and patients with the same type of cancer have often different outcomes. Polycomb proteins are emerging as important factors involved in breast cancer formation. Polycomb proteins play a crucial role in embryogenesis, early development, stem cell renewal and establishing and maintaining cell identity. Their alteration leads to mis-regulation of several important cellular factors including tumour suppressors, DNA repair factors, cell cycle regulation factors and cell-cell interaction factors. In this thesis the importance of several polycomb proteins in breast cancer has been investigated. The effect of EZH2 knockdown has been tested in breast cancer cell lines expressing different level of the protein and with different features. The results obtained are in line with other studies and suggest that the effect of EZH2 down-regulation in breast cancer cells is dependent on cellular context. In vitro experiments, using both established breast cell lines and primary epithelial cells have been used for investigating the importance of CBX8 in breast cancer. The results obtained showed that the polycomb proteins CBX8 does not play a central role in malignant transformation of the mammary epithelial cells tested.

Screening of breast cancer patients for their tumour's prognostic marker status is necessary in determining the most suitable course of treatment. This is particularly important in the assessment of HER-2 expression status in identifying candidates who may respond to trastuzumab therapy. Current methods are limited in their effectiveness in accurately determining actual marker status.Discussed herein is an investigation into the development of a titanium dioxide nanoparticle system which may be applied as a medical imaging methodology through the use of positron emission tomography to gauge accurately a patient's HER-2 expression status in identifying candidates for trastuzumab therapy. The initial synthesis of organically coated ultra-small titanium dioxide nanoparticles is discussed in depth with respect to a range of coating molecules and further functionalisation. Additionally, methodology to elicit an exchange of these coat molecules is explored in detail resulting in the generation of TiO2 nanoparticles capable of forming long-term stable suspensions in water. An exploration of the synthesis of fluoride accepting groups for use in generating radiolabelled compounds is explored both successfully and unsuccessfully leading to the development of conditions suitable for radiolabelling aryltrifluoroborate compounds. Attempts to then combine these radionuclide accepting groups with biologically compatible TiO2 nanoparticles are discussed as an initial step toward the generation of a potential PET tracer. However, while this conjugation was achieved, a successful demonstration of the radiolabelling was not achieved requiring further focus on modulating the nanoparticle to easily allow its recovery from such reactions. Finally, an investigation into the effects of trastuzumab and cetuximab on FDG uptake by cells in vitro is discussed with respect to the potential of monitoring disease response to these drugs with conventional FDG-PET.

Each year more than 44,000 women are diagnosed with breast cancer and more than 12,000 of these die from the disease. These mortality rates are relatively high compared to the rest of Europe (Berrino et al., 2007; Mayor, 2003), which in part is due to the advanced stage of the disease at first presentation (McCready et al., 2005; Sant et al., 2003). Regular breast self-examination (BSE) can facilitate early detection (Cancer Research UK, 2008), yet it is not widespread. Identifying the motivational and contextual factors is likely to lead to the development of effective interventions as part of being ‘breast-aware’. This is particularly important given that mammography screening may not be effective (Crossing & Mansezewicz, 2003) or recommended for younger women (Cancer Research UK, 2008) and that BSE may be beneficial in offering women the opportunity to create a positive relationship with their body. This study considered the utility of a proposed Extended Health Belief Model (E-HBM) as a framework for understanding women’s knowledge, beliefs and behaviour. It sought to develop a new way to provide guidance and to encourage BSE by investigating the effect of autobiographical accounts of breast-cancer patients and a multimedia BSE support programme comparing video-enhanced or static guided instructions. The study adopted a 2x2 mixed methods design and 60 white British women aged between 19 and 67 participated. Analysis of Covariance revealed an interaction effect of autobiographical accounts and BSE support on BSE frequency and proficiency and regression analyses examined the utility of the E-HBM, with confidence being the main predictor. Moreover, thematic analysis elicited five themes; Previous Experience, BSE Irregularity, Perceived Susceptibility, Coping Style and The Usability of the 5 Step Model of BSE. The study concludes that the 5-step technique encourages BSE through creating a more pleasant experience, as women form an improved relationship with their breasts. Both the video-enhanced and static BSE supports are effective in terms of encouraging BSE and the effect of autobiographical accounts appears to be dependent on the type support. The practical implications and direction for future BSE interventions are discussed.

Breast cancer is the single most prevalent form of cancer in the UK today, accounting for around 16% of all diagnoses, and around 31% of diagnoses in women. The survival rates are good, however the prognosis is heavily dependent on the stage to which the cancer has progressed at diagnosis. In order to help accurately determine this stage, the sentinel lymph node of patients undergoing tumour resection surgery is removed and examined cytologically for the presence of cancerous cells. This examination of the lymph node is currently the rate-limiting step in the operation as a whole. There is evidence in the literature to suggest that cancerous tissue has a measurably different infrared spectrum from healthy tissue owing to chemical and morphological differences in the tissue. There is further evidence to suggest that in the visible and near infrared region, the spectra of healthy lymph node tissue is different from that of cancerous tissue. This thesis details a project, performed in collaboration with a surgical team at St Mary's Hospital, Newport, Isle of Wight, to analyse spectral images taken in the visible and near infrared, of biopsied lymph node tissue. In the course of the project, an unsupervised classificaton technique, based on an extension to the well establised 'spectral angle', was developed to analyse the spectral images. Psoriasis affects 2-3% of the UK population causing itchy and/or painful plaques on the skin. One of the main treatments for psoriasis is UV phototherapy, exposure to which is a risk factor for burning and the development of cancers. This thesis details an investigation into the possibility of developing a targeted UV phototherapy system based on spectral imaging to delineate the plaques and a proposed new UV laser for treatment.