One of the best ways to diagnose breast cancer early or to predict therapeutic response is to use serum biomarkers. Unfortunately, for breast cancer, we do not have effective serological biomarkers. We hypothesized that novel candidate tumor markers for breast cancer may be secreted or shed proteins that can be detected in tissue culture supernatants of human breast cancer cell lines. A two-dimensional liquid chromatography-tandem mass spectrometry (2D-LC-MS/MS) strategy was utilized to identify and compare levels of extracellular and membrane-bound proteins in the conditioned media. Proteomic analysis of the media identified in excess of 600, 500 and 700 proteins in MCF-10A, BT474 and MDA-MB-468, respectively. We successfully identified the internal control proteins, kallikreins 5, 6 and 10 (ranging in concentration from 2-50 µg/L), as validated by ELISA and confidently identified HER-2/neu in BT474 cells. Sub-cellular localization was determined based on Genome Ontology (GO) for the 1,139 proteins, of which 34% were classified as extracellular and membrane-bound. Tissue specificity, functional classifications and label-free quantification were performed. The levels of eleven promising molecules were measured in biological samples to determine its discriminatory ability for control versus cases. This screen yielded activated leukocyte cell adhesion molecule (ALCAM) as a promising candidate. The levels of ALCAM, in addition to the classical breast cancer tumor markers carbohydrate antigen 15-3 (CA 15-3) and carcinoembryonic antigen (CEA) were examined in 300 serum samples by quantitative ELISA. All three biomarkers effectively separated cancer from non-cancer groups. ALCAM, with area under the curve (AUC) of 0.78 [95% CI: 0.73, 0.84] outperformed CA15-3 (AUC= 0.70 [95% CI: 0.64, 0.76]) and CEA (AUC= 0.63 [95% CI: 0.56, 0.70]). The incremental values of AUC for ALCAM over that for CA15-3 were statistically significant (Delong test, p <0.05). Serum ALCAM appears to be a new biomarker for breast cancer and may have value for disease diagnosis.

<p>A growing number of studies have investigated the role of exercise both during and after a breast cancer diagnosis. Observational data suggests that regular endurance exercise is associated with a 20-50% reduction in cancer-specific mortality in women diagnosed with early stage breast cancer, compared to inactive women; however it is unclear whether there is a differential association across breast cancer subtypes. As a pre-requisite to guide future large phase II/III clinical trials, there is a critical need to confirm the biological plausibility of the exercise association in breast cancer patients as well as elucidate the underlying mechanisms of action via utilization of preclinical models. </p><p>In the present study we investigated the systemic effects of prescribed aerobic training in cancer patients and the direct impact on breast cancer cell subtype phenotypes. In order to test the in vivo significance, we interrogated aerobic training effects on breast cancer progression and tumor biology using syngeneic breast cancer mouse models. </p><p>Our results suggest that aerobic training may alter the host availability of pro-inflammatory and growth factor cytokines in patients with solid tumors. Modulation of systemic effectors in breast cancer patients compared to controls causes a differential phenotypic response on breast cancer cell subtypes. In vivo, aerobic training has a differential response on breast tumor progression compared to controls that is mediated by Hif1-&#945; and metabolic reprogramming of breast cancer cells.</p> / Dissertation

The presence of distributed micro-calcifications can be an indicator of early breast cancer. On the mammogram, they appear as bright smooth particles superimposed on the normal breast image background. Radiologists determine the occurrence of this lesion by detecting the individual micro-calcifications and then examining their distribution within the breast tissue. Due to the visual complexity of the mammogram, the detection sensitivity is usually less than 100%. The digital environment has the potential to increase the radiologist's accuracy. We have developed a computer aided detection (CAD) scheme that can identify clinically indicative clusters of micro-calcifications. The CAD algorithm emulates some aspects of the radiologists' approach by using contrast texture energy segmentation and morphological distribution analysis. On a local database of 61 mammograms digitised at 100μm with 8 bits intensity resolution, the CAD returns: a) 85% sensitivity (91% for malignant lesions and 78% for those that are benign), b) 0.33 false positive clusters (FPC) per image and c) 92% specificity. Therefore, the output from the CAD is shown to compare favourably with the performance of an expert radiologist. It also compares favourably with other CAD techniques, exceeding many algorithms which employ a higher level of mathematical complexity. The scheme is tested on an international database provided by the Mammographic Image Analysis Society. In this case it returns a) 96.4% sensitivity (100% for malignant lesions and 92% for those that are benign) b) 2.35 FPC rate per image and c) 33% specificity. The higher FPC rate is attributed to the different acquisition and production of the digital mammograms. It is concluded that this can be reduced by employing a shape analysis procedure to the CAD's final output. It is shown that the image processing principles we have implemented are generally successful on databases which are produced at other centres under different technical conditions.

Globally, breast cancer is the most abundant cancer form in women, in Sweden about 20 women are diagnosed with breast cancer every day .Interactions between genetic and external factors are the contributing factors while metastasis formation is the leading cause of death. Cancer is in need of vessels,to get the nutrients and oxygen it needs in order to survive. Therefore,the aim of the study is to analyze and compare the groups of high and low differential cancer vessels of the respective form, and to see if any type contained more vessels than the other. The study is based on 20 invasive ductal breast cancer samples, ten of them were high differentiated and the other ten were low differentiated. To assess the number of vessels, immunhistochemical staining with CD31 antibody was performedCD31 is an adhesion molecule present on endothelial cells. The group of low differentiated gradebreast cancer tissue had significantly more vessels compared with the high differentiated breast cancer tissues. To prove these test results, more cancers must be analyzed.

The activity of S-adenosyl-L-methionine Decarboxylase (E. C. 4.1.1.50) was measured in two different normal and five different tumorous mouse mananary tissue samples. The soluble fraction was assayed for SAM decarboxylase activity using 14COOHlabeled SAM as the substrate. The amount of 14002 evolved was measured using a liquid scintillation counter.There was a significant elevation of SAM decarboxylase activity in the tumor sample group. The activities of the tumor tissue was x = 15.62 and SD = 8.24. The activity in the normal samples was too low to measure:These results suggest an increase in not only SAM decarboxylase activity, but also in the levels of spermidine and spermine. These increases are characteristics of cancer cells and may result in changes in enzyme activities and membrane phospholipid composition.

This study was carried out to investigate the fatty acid composition of tumoral, marginal and normal breast tissue in female breast cancer patients. Patients were recruited from St. Boniface General Hospital. A pre-operative blood sample was drawn. After surgery, sections were obtained from tumoral, marginal and normal breast tissues for histology and biochemical analysis. Extracted lipids from marginal tissue were significantly higher than those in normal or tumoral tissue. The lipid profile in tumoral tissue was significantly different in terms of fatty acid composition compared to normal and marginal tissue with less linoleic and alpha linolenic acid and more long chain polyunsaturated fatty acid of omega-3 and omega-6 series. Marginal tissue showed significantly less alpha linolenic acid compared to normal tissue. An inverse correlation existed between plasma level of 22:6 n-3 and breast cancer stage. We found different lipid profile in tumoral tissue compared to normal and marginal tissue.

Despite ongoing advancement in detection and treatment, breast cancer remains a major clinical challenge worldwide. Cancer has traditionally been conceptualized as a ‘disease of the genes’ by virtue of the mutagenic events necessary for its inception. It is now clear, however, that complex interactions take place between cancer cells and the array of non-cancerous cells and molecules in their immediate surroundings, known generally as the tumour microenvironment. Cancer-microenvironment interactions are increasingly recognized as processes that critically influence the outcome of disease. Cells of the host immune system are major components of the breast tumour microenvironment. While their presence in tumours is thought to reflect an attempt at disease eradication or containment, cancer cells can exploit the immune system through a variety of means, including the recognition of leukocyte-derived cytokines. As such, intratumoral leukocytes and high cytokine content are frequently associated with aggressive subtypes of breast cancer and poor prognosis. This dissertation explores the influence of one such cytokine, oncostatin-M (OSM), on the behaviour of breast cancer cells. Our results collectively demonstrate that OSM can rapidly and potently induce aggressive features in well-characterized cell models of luminal, well-differentiated breast cancer. These features include suppression of the important biomarker estrogen receptor-α (the key molecular target of endocrine therapy), gain of the breast cancer oncogene S100A7, loss of luminal epithelial differentiation and gain of mesenchymal features, and induction of a phenotype consistent with breast cancer stem cells. Each of these changes can potentially influence treatment responsiveness, the metastatic process, or both. Along with high levels of intratumoural leukocytes, the OSM-induced features listed above are known to associate with one another in human breast cancer. Tumours that display such characteristics have a poor prognosis and present the greatest challenges for modern breast cancer therapy, both because they are inherently prone to rapid metastasis and because targeted therapies for such tumours are lacking. The etiology of these aggressive disease subsets is largely unknown, and resolution of this issue would represent a major advancement in our understanding of breast cancer. Importantly, we found that expression of OSM and/or its receptor OSMR was reproducibly associated with these features in multiple breast cancer cohorts, largely confirming our experimental results. OSMR, in particular, was associated with poor clinical outcome. OSM signalling may thus provide a novel mechanistic explanation for the development of aggressive forms of breast cancer. If our findings are validated and expanded upon in future studies, OSM signalling could serve as a novel therapeutic target and may be an important consideration in the design and deployment of breast cancer immunotherapies. / Graduate

This thesis describes research carried out to explore the needs, expectations and experiences of the increasing number of women with a family history of breast cancer who present to primary care. This work was intended to inform clinical practice and policy, and to directly address women's needs where possible. Although a great deal of research has looked at the experiences of women with a family history of breast cancer in a specialist setting, when this work began, no research had yet been published on women's needs in primary care. This is particularly important, as general practitioners (GPs) are the first port of call, and often the main source of information, advice and support. A qualitative interview study was used for the preliminary work exploring women's subjective experiences of consulting primary care about a family history of breast cancer and understanding their primary care consultation needs (Chapter 2). This work was further elaborated upon using a prospective descriptive study to quantify the extent to which women shared the same views or experiences, and therefore, would benefit from certain changes in health care provision (Chapter 3). The qualitative and quantitative research showed that women's main primary care consultation needs were to discuss their risks of breast cancer with their GP and to receive verbal as well as take-home information. When this work was being carried out, no patient information was available suitable for a general population of women with breast cancer in the family who present to primary care. For this reason, a leaflet was developed based on women's information needs and the best available evidence (Chapter 4). The leaflet entitled Breast and/or Ovarian Cancer in the Family: Learning More about Your Risks and Options was evaluated with almost 200 women to ensure that it met their needs (Chapter 5). Over 90% of women were glad to have received the leaflet and felt that it provided the information they wanted to know. The implications of the work described in this thesis are that GPs could greatly assist their patients by acknowledging family history concerns as a legitimate reason for presenting to primary care, by providing verbal and take-home information and by inviting patients to return for future discussions if needed. Nationally accepted management guidelines for breast cancer family history and accompanying educational materials for use in primary care will also be instrumental in meeting patient needs and promoting informed choice in this new and difficult area of medicine.

No description available.

The BCA2 E3 ligase is expressed in a majority of invasive breast cancers. BCA2 has inherent autoubiquitination activity which contributes to cell migration and proliferation processes. Here, ten novel BCA2 binding proteins were found using yeast and bacterial screening. Two of which were human homolog of Rad23 variant A (hHR23a) and 14-3-3σ. In vivo and in vitro assays confirmed that both hHR23a and 14-3-3σ bound BCA2 and were co-expressed with BCA2 in breast cancer cells. Interaction of BCA2 with hHR23a and 14-3-3σ affect the autoubiquitination and auto-degradation activity of BCA2. Multi-ubiquitination of hHR23a-bound BCA2 was dramatically lower than that of free BCA2, this corresponded to increased BCA2 expression and half-life. Furthermore, phosphorylated BCA2 protein was stabilized by interaction with 14-3-3σ, via substrate inhibition of BCA2 autoubiquitination. High expression of BCA2 is correlated with grade in breast cancer and regulation of this E3 ligase’s activity may be important to cancer progression.