The prognostic role of the non-canonical nuclear factor-kappa B pathway in renal cell carcinoma patients

Background: In the United Kingdom, 8,000 cases of renal cancer are diagnosed each year, with a 5-year survival rate of 50%. Treatment options are limited; a potential therapeutic target is the non-canonical nuclear factor-kappa B (NF-κB) pathway. This pathway plays a role in multiple oncogenic processes in solid tumors. The aim of this study was to investigate the non-canonical nuclear factor pathway in renal cell carcinoma (RCC). Materials and Methods: NIK, IKKα, and RelB were investigated via immunohistochemistry in a cohort of 192 patients with clear cell renal cancer. Results: High cytoplasmic NIK was associated with poorer cancer-specific survival (p = 0.006) and 10-year survival stratified from 85% (low) to 65% (high, p = 0.005). Similarly, high cytoplasmic RelB was associated with poorer cancer-specific survival (p = 0.041) and 10-year survival stratified from 88% (low) to 73% (high, p = 0.030). When clinicopathological characteristics were assessed, cytoplasmic NIK was associated with survival (p = 0.014), whereas cytoplasmic RelB was associated with increased tumor grade (p = 0.020) and decreased inflammation (p = 0.019). Upon multivariate analysis, it was found that cytoplasmic NIK was independently associated with cancerspecific survival (p = 0.009). Conclusions: The non-canonical NF-κB pathway is associated with poorer cancer-specific survival in RCC patients, making it a viable target for therapeutic intervention. Furthermore, cytoplasmic NIK is a potential prognostic biomarker for this disease

In reply to "Hynes S.O. et al. Back to the future: routine morphological assessment of the tumour microenvironment is prognostic in stage II/III colon cancer in a large population-based study"

No abstract available

The role of gamma delta T lymphocytes in breast cancer: a review

Gammadelta T (γδT) lymphocytes have provoked interest in oncology, particularly as regards their potential use in immunotherapy, because of their unique ability to recognise antigens without a requirement for major histocompatibility complex antigen presentation, and to quickly activate an anti-tumour response. However, work in some cancers has suggested that they also have pro-tumourigenic activity. Their role in breast cancer is unclear. This review outlines the evidence to date in in vitro studies, in vivo mouse models and in human studies regarding the role of γδT lymphocytes in breast cancer. We describe the seemingly opposing roles of the predominantly circulating Vγ9Vδ2 subtype, which can suppress tumour growth through direct cytotoxicity, induction of apoptosis and inhibition of angiogenesis, and the predominantly tumour-infiltrating γδ1 subtype which can promote tumour growth and spread through immunosuppressant effects. We summarise the evidence in breast cancer for the mechanisms of action of γδT lymphocytes and describe how factors in the tumour microenvironment may affect their function, polarising them towards a pro-tumourigenic, immune-suppressing role. We also describe the experience to date of γδT lymphocytes in immunotherapy for breast cancer and suggest the direction of work going forward, particularly as regards different breast cancer subtypes

Phosphorylation of androgen receptors at serine 515 is a potential prognostic marker for triple negative breast cancer

1.7 million cases of breast cancer are diagnosed every year with 522,000 deaths. Molecular classifications of breast cancer have resulted in improved treatments. However, treatments for triple negative breast cancer (TNBC) are lacking. Analysis of molecular targets for TNBC is a priority. One potential candidate is androgen receptor (AR) phosphorylation. This study assessed the role of AR phosphorylation at ser81/ser515 and their two upstream effectors, cyclin-dependent kinase 1 (pCDK1) and extracellular-regulated kinase 1/2 (pERK1/2) in 332 ductal breast cancer patients by immunohistochemistry. pERK1/2 combined with AR-515 associated with improved cancer-specific survival (CSS, p = 0.038), decreased size (p = 0.001), invasive grade (p < 0.001), necrosis (p = 0.003), b-lymphocytes (p = 0.020), molecular subtype (p < 0.001) and estrogen receptor (ER)/progesterone receptor (PR)-status (p < 0.001). The cohort was therefore stratified into ER+ve and ER-ve patients. In ER+ve tumours, pERK1/2 combined with AR-515 associated with improved CSS (p = 0.038), smaller size (p = 0.004), invasive grade (p = 0.001), decreased b-lymphocytes (p = 0.013) and increased plasma cells (p = 0.048). In contrast, in TNBC patients, phosphorylation of AR-515 associated with poorer CSS (p = 0.007). pERK1/2 combined with AR-515 associated with decreased inflammation (p = 0.003), increased tumour stroma (p = 0.003) and tumour budding (p = 0.011), with trends towards decrease CSS (p = 0.065) and macrophage levels (p = 0.093). In Conclusions, AR-515 may be an important regulator of inflammation in breast cancer potential via ERK1/2 phosphorylation. AR-515 is a potential prognostic marker and therapeutic target for TNBC

Immunotherapy: enhancement the efficacy of this promising therapeutic in multiple cancers

Cancer treatments often reach a refractory period leading to treatment failure and patients developing disease recurrence. This can be due to tumour cells escaping the immune response and creating an immunosuppressive microenvironment enhancing cancer progression. Immunotherapy has become a promising tool for cancer treatment as it restores the anti-tumour response of the patient’s immune system. Immune checkpoint inhibitors are the most widely studied immunotherapies worldwide and are now approved for multiple cancers. However, CAR-T cell therapy has also shown promise by targeting T-lymphocytes that are genetically modified ex vivo to expressed chimeric antigen receptors and this is now approved to treat some haematological cancers. Although immunotherapy has shown successful treatment outcomes in multiple cancers, some patients do not respond to this treatment. Therefore, approaches to enhance the efficacy of immunotherapies are likely to be the key to improving their effectiveness. Therefore, combination therapies of checkpoint inhibitors +/- chemotherapy are at the forefront of current research. Furthermore, biomarkers that predict treatment response are now beginning to emerge. Additionally, utilizing nanoparticles as a new-targeted drug delivery system to enhance CAR-T cell therapy may enhance the efficacy of the cells when re-infused within the patient. Even if efficacy is enhanced, severe immune-related adverse events (irAEs) occur that are life threatening and could lead to therapy being stopped. Therefore, predictive biomarkers for toxicity are also needed to improve both the patient’s quality of life and treatment outcomes. This review will look at the current immunotherapies in clinical trials and discuss how to enhance their efficacy

The effects of extracellular sodium chloride on the activity and expression of Na,K-ATPase in primary cultures of dogfish (Scyliorhinus canicula) rectal gland epithelial cells

Dogfish, Scyliorhinus canicula, rectal gland epithelial cells were successfully cultured using two different techniques: 1) a perfusion based technique and 2) a modified Valentich's technique. The morphology of the primary rectal gland epithelial cell cultures was investigated using light, fluorescence and electron microscopy. These studies demonstrated that the cell cultures express most of the structural features of native shark rectal gland cells, including numerous mitochondria, complex tight junctions and extensive membrane folding. The cultured cells using the perfusion technique adopted an extremely flattened morphology when grown on collagen. These cells whether grown in suspension or on collagen, displayed a striking level of vacuole formation, these vacuoles were not associated with transport epithelia. The rectal gland cell cultures were then used to investigate the effect increasing extracellular sodium chloride concentration has on rectal gland cells Na, K-ATPase activity. Increasing sodium chloride concentration in the growth medium by 50% (240 mM to 360 mM) resulted in a transient 3-4 fold increase in Na, K-ATPase activity in cell homogenates approximately 12 hours after the medium change. The response was dependent upon both sodium and chloride ions and was also inhibited by the loop diuretic bumetanide (0.1 mM within 30 minutes), indicating that entry of the ions into the cell is via the Na, K, C1 cotransporter. Incubation of cells in normal medium in the presence of the sodium ionophore monensin also resulted in a dose dependant sustained increase in Na, K-ATPase activity following a 12 hour incubation. The increase in Na, K-ATPase activity associated with increased extracellular sodium chloride concentration was only seen in cells grown on collagen and not in cells grown in suspension. Increases in activity are sensitive to the protein synthesis inhibitor cycloheximide (10 mug/ml), but not the transcriptional inhibitor actinomycin D suggesting that up-regulation of the Na, K-ATPase occurs at the level of translational regulation. Unfortunately this result could not be confirmed using Northern analysis due to unforeseen difficulties in extracting sufficient RNA from the cell cultures. Addition of bumetanide (0.1 mM) to cells grown in normal medium caused a rapid but reversible down-regulation (by 70%) of basal Na, K-ATPase activity within 30 minutes. The anti-microtubular agent colchicine (0.1 mug/ml) inhibited the bumetanide induced down-regulation of Na, K-ATPase and also the recovery of activity following bumetanide removal. The rectal gland cell cultures were used to investigate potential hormonal regulators of the shark rectal gland. The effect of the putative regulators of sodium chloride secretion scyliorhinin II and sCNP on intracellular concentrations of cAMP and cGMP was investigated. The cell cultures were shown be hormonally active as they responded with an increase in intracelllular cAMP concentration to forskolin, PGE1 and PGE2. When scyliorhinin II (10 muM) and IBMX (1 mM) was perfused through the isolated rectal gland a 2 fold increase in cAMP concentration was found in the perfusate after 8 minutes, however no increase was seen in cAMP levels when cell cultures were treated with scyliorhinin II. Shark CNP increased cGMP concentrations in the perfusates of the perfused rectal gland by up to four fold after seven minutes but there was no consistent effect on cGMP concentrations in the cultured cell monolayer. In conclusion it is believed that sCNP and scyliorhinin II mediate their actions on the regulation of sodium chloride secretion by the rectal gland at the vascular level, controlling the extent of perfusion of the gland. This study showed that high salt levels in the medium of shark rectal gland cell monolayers increased the measurable Na, K-ATPase activity and that this response was dependent on protein synthesis but not transcription. It also showed that the response is inhibited by the loop diuretic bumetanide, indicating that entry of the ions into the cell is via the Na, K, Cl cotransporter and that the increase in Na, K-ATPase activity is presumably due to an increase in intracellular sodium concentration. The hormones sCNP and scyliorhinin II appear to mediate their actions on the regulation of sodium chloride secretion by the rectal gland at the vascular level controlling the extent of perfusion of the gland. In conclusion although sodium chloride transport in the dogfish rectal gland requires much more investigation, this study has hopefully proved that dogfish epithelial cell cultures provide a good model for further investigations involving the regulation of activity and expression of the sodium pump

Loss of signal transducer and activator of transcription 1 is associated with prostate cancer recurrence

STAT1 loss has previously been implicated in cell line studies to modify prostate cancer cell growth and survival, however the clinical significance of this has not previously been established. This study investigated if STAT1 loss was associated with patient outcome measures and the phenotypic consequence of STAT1 silencing. STAT1 expression was assessed in two patient cohorts with localised (n = 78) and advanced prostate cancer at initial diagnosis (n = 39) by immunohistochemistry (IHC). Impact of STAT1 silencing on prostate cancer cells lines was assessed using Cell Death detection ELISA, TLDA gene signature apoptosis arrays, WST-1 assay, xCELLigence system, clonogenic assay, and wound healing assay. In the localised patient cohort, low expression of STAT1 was associated with shorter time to disease recurrence (3.8 vs 7.3 years, P = 0.02) and disease specific survival (6.6 vs 9.3 years, P = 0.05). In the advanced patient cohort, low expression was associated with shorter time to disease recurrence (2.0 vs 3.9 years, P = 0.001). When STAT1 was silenced in PC3 cells (AR negative) and LNCaP cells (AR positive) silencing did not influence levels of apoptosis in either cell line and had little effect on cell viability in the LNCaP cells. In contrast, STAT1 silencing in the PC3 cells resulted in a pronounced increase in cell viability (WST-1 assay: mock silenced vs STAT1 silenced, P < 0.001), clonagenicity (clonogenic assay: mock silenced vs STAT1 silenced, P < 0.001), and migration (wound healing: mock silenced vs STAT1 silenced, P < 0.001). In conclusion, loss of STAT1 may promote prostate cancer recurrence in AR negative patients via increasing cell viability

Literacy practices in the learning careers of childcare students

This paper draws from the Literacies for Learning in Further Education research project, funded through the Teaching and Learning Research Programme. Drawing on the empirical study of literacy practices in eight Childcare courses in Scotland and England, we seek to demonstrate that, integral to the learning careers of students are literacy careers through which their learning is mediated. In the process, by drawing upon the lens of literacy, we also challenge some of the common sense understandings of learning in childcare. In particular we suggest that the literacy practices of lower level courses can be more diverse than those of higher level courses, producing confusing literacy careers for the students involved. We also point to the complexity of the literacy careers in childcare, given that students are required to mediate different aspects of their experience through literacy. In particular there are the mediations made possible by the use of information technology and those entailed in relating work placements to classroom practice. We argue that students on vocational courses have complex literacy careers and that a literacies approach to learning helps to reveal this complexity