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Designs and Samples. Four questionnaire-based studies were conducted: one prospective study with 67 adults, followed up at three months; and three cross-sectional studies with sample sizes of 136, 147 and 283. Each study included female and male participants, who were aged 20 or over and lived in Great Britain. Measures. A range of individual differences were measured within the four studies. These included health locus of control beliefs, coping responses, general anxiety, anxiety in relation to health and to performing self-examinations, worry about breast or testicular cancer, perceived breast or testicular cancer risk, dispositional optimism, health optimism, and socially desirable responding. In addition, female participants provided self-reports of their breast self-examination behaviour and in the final study, whether or not they were ‘breast aware’; while male participants gave self-reports of their testicular self-examination behaviour. Results. Multiple regression analyses revealed that worry about breast or testicular cancer and anxiety about performing self-examinations operated as the most consistent independent predictors of self-examination behaviour. In addition, ANCOVAs showed that perceived breast or testicular cancer risk and dispositional optimism were significantly associated with self-examination frequency. Across the four studies, associations between the individual differences and adults’ self-examination behaviour were generally evidenced as being similar for females and males. Utilising data from the final study, a model of individual differences and breast or testicular self-examination behaviour was constructed and proposed. Conclusions. The present research has demonstrated associations between a number of individual differences and breast or testicular self-examination. These findings indicate the potential impact of individual differences on adults’ self-examination behaviour.

Background and Aims: Breast cancer is the second most common cause of cancer deaths in women. It is a tumour which has been extensively studied at a molecular level and, compared to other solid tissue tumours, our understanding of its biology is extensive. There are however some patients who are considered to have good prognostic feature of their tumours who go on to die from their disease. Transcription factors are the end point of many cell signalling pathways. They form the link between exogenous hormones and growth factors and DNA transcription. For the purpose of this study 3 different transcription factors have been selected for investigation. STAT3 is activated by various growth factors and cytokines including EGF. It is classified as an oncoprotein as its activation can mediate tumorgenisis in nude mice. STAT3 has been shown to confer resistance to apoptosis in breast cancer cells and it is associated with poor outcome in high risk breast cancers. SP1 is a transcription factor which is essential in the expression and the action of estrogen receptors (ER). It is known to be over expressed in other solid tissue tumours but there has been little work into its role in breast cancer. NFkB is activated in many cell survival settings. It is involved in the transcription of anti-apoptotic genes and also plays a role in cell proliferation, angiogenisis and cell adhesion. It is associated in breast cancers with an over expression of the oncogene Bcl-2. It has not been show to be a marker of prognosis but does appear to identify breast cancers with a poor response to chemotherapy. The aim of this study is to investigate the role of these transcription factors in the behaviour of breast cancers and the outcome of the disease. It will also investigate the affect of EGF and estogen stimulation on STAT3 activation in breast cancer cell lines. Methods: This study consists of 2 elements. Firstly an assessment of transcription factor expression in breast cancer samples and secondly a cell model experiment to investigate the stimulation of STAT3 activation. A cohort of 213 patients who presented to the Queen Elizabeth Hospital with invasive breast cancer in 1999 was selected. Tumour samples from these patients were retrieved and using immunohistochemistry were tested for the expression of STAT3, SP1 and NFkB. These results were then correlated with pathological features of the tumours, tumour receptor status (ER, PR HER2 and EGFR) and outcome of the disease. Two cell lines, MCF7 and SKBr3, were cultured in depleted medium. These cells were then stimulated with estrogen and EGF alone and in combination. Flow-cytometry was then used to quantify the levels of phosphorylated STAT3 in the 2 cell lines over a 3 day time course. The level of phosphorylation was then compared to the control lines to asses the effect of stimulation. Results: 209 breast cancers were successfully analysed for the expression of STAT3, 27% of these cancers expressed nuclear STAT3. The results demonstrated a significant correlation of STAT3 expression with cancers of a high grade (p=<0.001), increasing tumour size (p=0.004), vessel space invasion (p=0.034) and lymph node metastases (p=0.015). STAT3 expression was shown to be significantly correlated to high Nottingham prognostic index (NPI) scores. With regards to receptor status it was show that STAT3 expression was significantly associated with ER negative and PR negative cancers (p=0.003), whereas there was no relationship with HER2 status. The results did show that there was a significant relationship between STAT3 expression and EGFR positive cancers (p=0.007). When disease outcome was investigated it was shown that there was a trend towards improved survival in the STAT3 negative group and a significant relationship between STAT3 expression and disease recurrence at 5 years (p=0.04). SP1 expression was determined in 208 of the cancer samples with 33% of the tumours having strong nuclear staining. There was no significant relationship between SP1 expression and any of the pathological features mentioned. SP1 expression was related to ER positive tumours (p=0.015). Though there was no relationship with 5 year survival it appears that SP1 expression does reduce the risk of late (>2yr) disease recurrence (p=0.005). NFkB was over expressed in 15% of the 208 cancers samples. Again a significant correlation was shown with high grade tumours (p=0.001) and large tumours (p=0.014). NFkB expression was also shown to be more prevalent in ER negative cancers (p=0.006) and EGFR positive tumours (p=0.007). There was no significant relationship between NFkB expression and disease outcome. The cell model results showed that in the EGFR positive ER negative cell line (SKBr3), EGF stimulation resulted in a biphasic response of STAT3 phosphorylation, whereas estrogen had no effect on phosphorylation. In the ER positive MCF7 cells, which express low levels of EGFR, again EGF stimulation resulted in a biphasic response curve. Estrogen stimulation does cause an increase in activation but when estrogen is added to EGF stimulation there is an inhibition of STAT3 phosphorylation. Conclusions: This study has demonstrated that STAT3 and SP1 expression is important in disease outcome in breast cancer patients. Though there are differences in levels of expression, NFkB does not appear to have a role in breast cancer outcome. The cell model has show that EGF stimulation of EGFR positive cell lines results in increased STAT3 activation and also that this effect is inhibited by the addition of estrogen stimulation. These results raise important questions which are discussed in the study and suggest areas for further investigation.

The FOXP3 transcription factor can regulate T cell migration by inhibiting expression of CXCR4, the receptor for the chemokine CXCL12. The increased expression of CXCR4 by breast cancer cells can drive metastatic migration towards sites that express CXCL12. Intracellular trafficking of FOXP3 to the nucleus is required in order for this factor to function. The presence of alternative splicing forms, mutations and post-translationally modified forms may disrupt FOXP3 nuclear localization. We hypothesised that FOXP3 tumour suppressor is inactive in breast cancer causing an increase in CXCR4 expression and the development of metastasis. The expression of FOXP3 and CXCR4 were measured at mRNA and protein (immunohistochemistry, immunofluorescence, FACS) levels. FOXP3 DNA sequences of normal and cancer cells were analysed. Stable FOXP3 overexpressing breast cancer transfectants were used to investigate the potential of FOXP3 to regulate chemotaxis. Normal breast epithelial cells (both patient-derived tissues and laboratory cultured cell lines) expressed FOXP3 in their nuclei but did not express CXCR4. Breast cancer cells overexpressed CXCR4 (p<0.05), whereas FOXP3 expression was decreased (p<0.05) and confined to the cytoplasm with negligible nuclear expression. Metastases expressed less FOXP3 and more CXCR4 than primary cancers (p<0.05). FOXP3 sequencing in breast cancer cell lines did not reveal mutations. However, there were at least three bands on the PCR electrophoreses gel. The predominant form in breast cancer cells contained an insertion of 120bp within the forkhead domain. Transfection of breast cancer cells with wild-type FOXP3 restored its nuclear expression, reduced CXCR4 expression and inhibited cell migration. This study demonstrated failure of nuclear localisation of FOXP3 in breast cancer cells and an inverse correlation between this failure and CXCR4 expression. Disruption of FOXP3 nuclear localisation may be due to abnormal co-expression of FOXP3 splice variants leading to increased CXCR4 expression and acquisition of the capacity to metastasize.

The aim of this study was to elucidate the underlying molecular genetic mechanisms of radiation-induced breast carcinogenesis. Younger women are more at risk of developing breast cancer than older women following radiation exposure. Circulating oestrogen levels are highest during adolescence and early adulthood and oestrogen has a known transforming effect on breast epithelial cells. One hypothesis suggests that radiation and oestrogen synergise to drive breast cell transformation. There are currently no known genetic markers of radiogenic breast cancer. In order to investigate genetic alterations associated with radiogenic breast cancer in vitro models of radiation-induced breast epithelial cell transformation was developed. The immortalised, non-transformed breast epithelial cell line MCF-10A was exposed to fractionated doses of X-rays in the presence or absence of additional oestrogen. Radiation-treated cells displayed several phenotypic changes some of which provided evidence of cell transformation, including loss of contact inhibition and change to mesenchyme cell morphology. Genomic analysis of radiation treated cells using high-density polymorphism arrays identified a gene deletion of the POU2F1 transcription factor and amplification of the c-MYC proto-oncogene. POU2F1 has a role in cellular stress response and mediates DNA damage response through interactions with BRCA1. Amplification of c-MYC has previously been identified in breast cancers of survivors of the atomic bombs during World War II. Genetic alteration in POU2F1 and c-MYC may therefore be linked to radiation-induced breast cell transformation. Changes in gene copy-number were confirmed by fluorescent in situ hybridisation (FISH) and alterations in protein expression by western analysis. Gene copy number and expression of POU2F1 and c-MYC was investigated in a cohort of radiogenic and sporadic breast cancer tissue samples by FISH analysis and immunohistochemistry. Expression of c-MYC was higher in radiation-induced breast cancers compared to sporadic breast cancers (p = 0.002), as was the mean number of copies of c-MYC (p = 0.030). Loss of expression of POU2F1 was identified in one of 18 radiation-induced breast cancers but was not observed in sporadic breast disease (0/33). In summary, a cell model of radiation-induced breast cell transformation identified POU2F1 deletion and c-MYC amplification as putative markers of transformation, which were subsequently identified in primary tissue samples, suggesting a role for these alterations in the development of radiogenic breast cancer.

Breast cancer is the most common form of cancer in women of all ethnic groups. The diagnosis and treatment of the disease can be challenging and distressing for any individual. Research in this area has documented a range of physical, psychological and social consequences. However, very little work within the existing literature captures the experiences of Black and Minority Ethnic (BME) women, especially in the UK. The limited available research, based on American studies, show that while similarities in White and BME women’s breast cancer experiences exist, there are also aspects that are unique and specific to BME women, which are influenced by socio-cultural norms, behaviours and beliefs. Therefore, the aim of this research was to explore the psychosocial impact of breast cancer diagnosis and treatment in British Black and South Asian women. To research this area, a qualitatively driven mixed method approach was adopted. Four studies were undertaken, 3 qualitative and 1 quantitative. The first study was exploratory in nature, in which semi-structured one-to-one interviews were carried out with 11 Black and 11 South Asian women. The findings of this study informed the subsequent studies, the second of which consisted of focus group interviews with 10 Black and 10 South Asian women to explore the ways in which breast cancer experiences are managed. The third study involved interviews with 5 South Asian Indian, Gujarati-speaking women (with limited English proficiency). The findings from these studies show that psychosocial and cultural factors (such as support, appearance concerns, healthcare experiences, cultural beliefs about cancer, understanding of cancer and language barriers) play an important role in shaping BME women’s experiences of breast cancer. These findings informed the development of the final questionnaire study. This study investigated the psychosocial impact of breast cancer and aimed to compare breast cancer experiences between White, Black and South Asian women. A total of 173 women participated (80 White, 40 Black and 53 South Asian). The results from this study show that psychological distress, social support and receiving chemotherapy treatment are associated with quality of life. Furthermore, similarities and differences in Black, South Asian and White women’s breast cancer experiences (in relation to psychological distress, quality of life, control beliefs (pertaining to cancer), body image concerns and sources of support) are evident. The findings from this research highlight important implications for policy makers, healthcare professionals and researchers to enhance awareness and understanding, and to ensure the provision of culturally competent care and support to future breast cancer patients.

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No description available.

Breast cancer presenting in women aged ≤ 35 years has a more aggressive behaviour, and thus poorer prognosis, which is thought to be due to differences in the tumour biology of these tumours compared to those from older women. The aim of this study was to examine the expression of 9 novel gene targets (A-kinase anchor protein-1 (AKAP1), Acidic protein rich in leucines (APRIL), CCAAT enhancer binding protein alpha (C/EBPα), Damage-specific DNA binding protein 2 (DDB2), Granulin, Nuclear Receptor Coactivator 3 (NCOA3), Retinoic acid receptor Responder 3 (RARRES3), Retinoblastoma binding protein 4 (RBBP4), and Transforming Growth Factor beta Induced (TGFβI) identified previously by a cDNA microarray in breast cancers and female controls by RT-qPCR, western blotting, and immunohistochemistry. Six breast cell lines, 9 samples of organoids from reduction mammoplasty tissues, and 35 tumour tissues (20 cases >35 years, 15 cases ≤35 years in age) were analysed for the expression of the nine target genes using real time quantitative RT-PCR. Of the nine target genes investigated, five showed differences between normal and cancers ≤35, or between breast cancers ≤35 and those >35 years. NCOA3 and RARRES3 showed elevated levels of mRNA in breast cancers ≤35 years compared to those >35 years (p= 0.001 and p=0.002 respectively). Compared to the normal breast, TGFβI showed a reduced level of mRNA expression in both younger and older cases (p= 0.026 and p=0.001 respectively), while DDB2 and C-EBPα showed a reduced level of mRNA expression in younger group only (p=0.002 and p=0.001 respectively). NCOA3 protein expression examination using western blotting found high levels in the ER+ve cell lines MCF-7, ZR-75-1 and T47-D with a weak expression in ER−ve cell lines HBL-100 and MDA-MB-468. RARRES3 protein expression was found in 4 breast cell lines (HBL-100, MDA-MB-468, MCF-7, and ZR-75-1). IHC found expression of NCOA3 in younger and older tumours including ER+ve and ER−ve cases. This study identifies NCAO3 and RARRES3 as potential markers for breast cancers in younger women, but the data need confirmation in a larger series of cases.

There is increasing evidence that stromal fibroblasts promote breast cancer invasion. This thesis aimed to dissect phenotypic and genotypic differences between fibroblasts from normal donors and patients with breast cancer and to relate these to their capacity to promote tumour cell invasion.;Immunohistochemical analysis demonstrated up-regulation of alpha4 integrin, alphavbeta3 integrin and alpha-SMA in peri-tumoural fibroblasts compared to normal fibroblasts in vivo.;Primary isolated peri-tumoural fibroblasts demonstrated greater invasion promoting capacity (IPC) than normal breast fibroblasts, however, matched non-breast (dermal) fibroblasts from patients with breast cancer also exhibited higher (IPC) than dermal fibroblasts from normal donors implying that intrinsic genetic differences exist.;Since fibroblast derived matrix metalloproteinases (MMP's) are known to contribute to tumour invasion, fibroblast IPC was related to expression levels and release of MMP's. No significant difference in MMP expression, as measured by real time PCR, was detected between normal and tumour donors. However, a trend towards higher levels of MMP release and IPC of fibroblasts populations was observed.;Functional promoter polymorphisms have been shown to influence expression of some MMPs. This thesis demonstrated a trend towards increased frequency of the high expression MMP-3 5A allele and both the tumour group and high IPC fibroblasts. In addition the 5A allele correlated with increased levels of MMP-3 release.;Furthermore, regulation of fibroblast IPC and MMP-3 release was shown to be co-ordinately regulated by alpha and alpha5 integrins.;In conclusion, this thesis demonstrates phenotypic differences between fibroblasts isolated from normal donors and patients with breast cancer which influence their capacity to promote tumour invasion. Also intrinsic genetic differences exist between patients which relate to both IPC and MMP release.