Iain Brown

OPCML at 11q25 is epigenetically inactivated and has tumor-suppressor function in epithelial ovarian cancer

Epithelial ovarian cancer (EOC), the leading cause of death from gynecological malignancy, is a poorly understood disease. The typically advanced presentation of EOC with loco-regional dissemination in the peritoneal cavity and the rare incidence of visceral metastases are hallmarks of the disease. These features relate to the biology of the disease, which is a principal determinant of outcome. EOC arises as a result of genetic alterations sustained by the ovarian surface epithelium (OSE; ref. 3). The causes of these changes are unknown but are manifest by activation of oncogenes and inactivation of tumor-suppressor genes (TSGs). Our analysis of loss of heterozygosity at 11q25 identified OPCML (also called OBCAM), a member of the IgLON family of immunoglobulin (Ig) domain–containing glycosylphosphatidylinositol (GPI)-anchored cell adhesion molecules, as a candidate TSG in EOC. OPCML is frequently somatically inactivated in EOC by allele loss and by CpG island methylation. OPCML has functional characteristics consistent with TSG properties both in vitro and in vivo. A somatic missense mutation from an individual with EOC shows clear evidence of loss of function. These findings suggest that OPCML is an excellent candidate for the 11q25 ovarian cancer TSG. This is the first description to our knowledge of the involvement of the IgLON family in cancer.

miRNA-34a is associated with docetaxel resistance in human breast cancer cells

Docetaxel is a chemotherapy drug to treat breast cancer, however as with many chemotherapeutic drugs resistance to docetaxel occurs in 50% of patients, and the underlying molecular mechanisms of drug resistance are not fully understood. Gene regulation through microRNAs (miRNA) has been shown to play an important role in cancer drug resistance. By directly targeting mRNA, miRNAs are able to inhibit genes that are necessary for signalling pathways or drug induced apoptosis rendering cells drug resistant. This study investigated the role of differential miRNA expression in two in vitro breast cancer cell line models (MCF-7, MDA-MB-231) of acquired docetaxel resistance. MiRNA microarray analysis identified 299 and 226 miRNAs altered in MCF-7 and MDA-MB-231 docetaxel-resistant cells, respectively. Docetaxel resistance was associated with increased expression of miR-34a and miR-141 and decreased expression of miR-7, miR-16, miR-30a, miR-125a-5p, miR-126. Computational target prediction revealed eight candidate genes targeted by these miRNAs. Quantitative PCR and western analysis confirmed decreased expression of two genes, BCL-2 and CCND1, in docetaxel-resistant cells, which are both targeted by miR-34a. Modulation of miR-34a expression was correlated with BCL-2 and cyclin D1 protein expression changes and a direct interaction of miR-34a with BCL-2 was shown by luciferase assay. Inhibition of miR-34a enhanced response to docetaxel in MCF-7 docetaxel-resistant cells, whereas overexpression of miR-34a conferred resistance in MCF-7 docetaxel-sensitive cells. This study is the first to show differences in miRNA expression, in particular, increased expression of miR-34a in an acquired model of docetaxel resistance in breast cancer. This serves as a mechanism of acquired docetaxel resistance in these cells, possibly through direct interactions with BCL-2 and CCND1, therefore presenting a potential therapeutic target for the treatment of docetaxel-resistant breast cancer.

Alterations of β-tubulin isotypes in breast cancer cells resistant to docetaxel

Docetaxel is one of the most active drugs used to treat breast cancer. The cellular target of docetaxel is the microtubule, specifically the β‐tubulin subunit, that comprises a series of isotypes and that can modulate function. This study has examined the role of alteration in β‐tubulin isotypes in vitro and has sequenced the β‐tubulin gene to determine if there were mutations, both of which may represent important mechanisms of acquired resistance to docetaxel. Breast cancer cells, MCF‐7 (oestrogen‐receptor positive) and MDA‐MB‐231, (oestrogen‐receptor negative) were made resistant to docetaxel in vitro. Expression of β‐tubulin isotypes (class I, II, III, IVa, IVb, and VI) was determined at the RNA and protein level using RT‐PCR and western analysis, respectively. DNA sequencing evaluated the β‐tubulin gene. At the mRNA level, class I, II, III, and IVa β‐tubulin mRNA isotypes were over‐expressed in docetaxel‐resistant MCF‐7 cells when compared with the docetaxel‐sensitive parental cells. However, class VI β‐tubulin mRNA isotype expression was decreased in resistant cells. In MDA‐MB‐231 cells, there was a decrease in expression of the class I and class IVa β‐tubulin mRNA. However, there were increased expressions in class II, IVb, and VI β‐tubulin mRNA isotypes in resistant cells. Western analysis has confirmed corresponding increases in β‐tubulin protein levels in MCF‐7 cells. However, in MDA‐MB‐231 cells, there were decreased protein levels for class II and class III β‐tubulin. This study demonstrates that altered expression of mRNA β‐tubulin isotypes and modulation of β‐tubulin protein levels are associated with acquired docetaxel resistance in breast cancer cells. This allows further understanding and elucidation of mechanisms involved in resistance to docetaxel.

Plant Phenolics in the Prevention and Treatment of Cancer

Epidemiological studies indicate that populations consuming high levels of plant derived foods have low incidence rates of various cancers. Recent findings implicate a variety of phytochemicals, including phenolics, in these anticancer properties. Both monophenolic and polyphenolic compounds from a large variety of plant foods, spices and beverages have been shown to inhibit or attenuate the initiation, progression and spread of cancers in cells in vitro and in animals in vivo. The cellular mechanisms that phenolics modulate to elicit these anticancer effects are multi-faceted and include regulation of growth factor-receptor interactions and cell signaling cascades, including kinases and transcription factors, that determine the expression of genes involved in cell cycle arrest, cell survival and apoptosis or programmed cell death. A major focus has been the inhibitory effects of phenolics on the stress-activated NF-KB and AP-1 signal cascades in cancer cells which are regarded as major therapeutic targets. Phenolics can enhance the bodys immune system to recognize and destroy cancer cells as well as inhibiting the development of new blood vessels (angiogenesis) that is necessary for tumour growth. They also attenuate adhesiveness and invasiveness of cancer cells thereby reducing their metastatic potential. Augmentation of the efficacy ofstandard chemo- and radiotherapeutic treatment regimes and the prevention of resistance to these agents is another important effect of plant phenolics that warrants further research. Plant phenolics appear to have both preventative and treatment potential in combating cancer and warrant further, in-depth research. It is interesting that these effects of plant phenolics on cancer inhibition resemble effects reported for specific fatty acids (omega-3 PUFA, conjugated linoleic acids). Although phenolic effects in cells in vitro and in animal models are generally positive, observations from the less numerous human interventions are less clear. This is surprising given the positive epidemiological data and may relate to mixed diets and synergistic interactions between compounds or the bioavailability of individual compounds. Much of the work in vitro with phenolic compounds has utilized concentrations higher than the amount that can be obtained from the diet suggesting a role of fortified, functional foods in cancer suppression.

Reduced expression of p27 is a novel mechanism of docetaxel resistance in breast cancer cells

IntroductionDocetaxel is one of the most effective chemotherapeutic agents in the treatment of breast cancer. Breast cancers can have an inherent or acquired resistance to docetaxel but the causes of this resistance remain unclear. However, apoptosis and cell cycle regulation are key mechanisms by which most chemotherapeutic agents exert their cytotoxic effects.MethodsWe created two docetaxel-resistant human breast cancer cell lines (MCF-7 and MDA-MB-231) and performed cDNA microarray analysis to identify candidate genes associated with docetaxel resistance. Gene expression changes were validated at the RNA and protein levels by reverse transcription PCR and western analysis, respectively.ResultsGene expression cDNA microarray analysis demonstrated reduced p27 expression in docetaxel-resistant breast cancer cells. Although p27 mRNA expression was found to be reduced only in MCF-7 docetaxel-resistant sublines (2.47-fold), reduced expression of p27 protein was noted in both MCF-7 and MDA-MB-231 docetaxel-resistant breast cancer cells (2.83-fold and 3.80-fold, respectively).ConclusionsThis study demonstrates that reduced expression of p27 is associated with acquired resistance to docetaxel in breast cancer cells. An understanding of the genes that are involved in resistance to chemotherapy may allow further development in modulating drug resistance, and may permit selection of those patients who are most likely to benefit from such therapies.

Simulation scenarios of spatio-temporal arrangement of crops at the landscape scale

The spatial and temporal arrangement of crops is a conspicuous feature of rural landscapes. It has been identified as an important factor in many environmental issues, such as the coexistence of genetically modified (GM) and non-GM crops, and the mitigation of soil erosion. This paper examines a scenario-based approach for rapid generation and screening of crop allocations that meet users constraints without requiring mechanistic modelling. LandSFACTS (Landscape Scale Functional Allocation of Crops Temporally and Spatially) is a software application specifically designed to simulate such crop arrangement scenarios, whilst ensuring both spatial and temporal coherence with regard to the initial constraints. The software uses an empirical approach to allocate crops to fields (polygons in vector format) over a sequence of years, using a stochastic process (Markov chains) and rule-based constraints. Crop rotations are represented by transition probabilities complemented by other temporal constraints such as return period or prohibited sequences. Further spatial and temporal constraints on crop arrangement can be applied through separation distances, yearly proportions, and the application of statistical tests. The software outputs a crop allocation solution with a crop for every field for every year, respecting all user-defined constraints; the range of potential solutions can then be explored through multiple model runs. Metrics based upon the difficulty of obtaining such an allocation from the initial constraints are also generated. A case study is provided to demonstrate the use of combined agronomic and environmental criteria for exploring GM crop coexistence at the landscape scale.

Modelling future landscape change on coastal floodplains using a rule-based GIS

Abstract A large-scale, strategic-level method has been developed to facilitate dialogue with policymakers on future coastal land-use management, in the context of climate change and the implications of sea-level rise for floodplain areas. Impacts from different management policies are explored through their influence on patterns of flooding and consequent landscape change. The method is based upon a series of transition rules linking frequency of inundation by the sea to changes in land use and land cover at key threshold values. A high-resolution digital elevation model derived from Interferometric Synthetic Aperture Radar (IfSAR) provides the topographic information required for relating sea-level rise to tidal and flood frequency limits. The rule-based approach allows a rapid evaluation of multiple policy options and critical assumptions can be made more transparent and responsive to end-users than with conventional modelling approaches. A case study example is provided from the North Norfolk coast in eastern England, where the results imply that future sea-level rise has much broader policy implications for the human and ecological landscape than just flood risk to property.

Cannabinoids and omega-3/6 endocannabinoids as cell death and anticancer modulators

Cannabinoids-endocannaboids are possible preventatives of common diseases including cancers. Cannabinoid receptors (CB(½), TRPV1) are central components of the system. Many disease-ameliorating effects of cannabinoids-endocannabinoids are receptor mediated, but many are not, indicating non-CBR signaling pathways. Cannabinoids-endocannabinoids are anti-inflammatory, anti-proliferative, anti-invasive, anti-metastatic and pro-apoptotic in most cancers, in vitro and in vivo in animals. They signal through p38, MAPK, JUN, PI3, AKT, ceramide, caspases, MMPs, PPARs, VEGF, NF-κB, p8, CHOP, TRB3 and pro-apoptotic oncogenes (p53,p21 waf1/cip1) to induce cell cycle arrest, autophagy, apoptosis and tumour inhibition. Paradoxically they are pro-proliferative and anti-apoptotic in some cancers. Differences in receptor expression and concentrations of cannabinoids in cancer and immune cells can elicit anti- or pro-cancer effects through different signal cascades (p38MAPK or PI3/AKT). Similarities between effects of cannabinoids-endocannabinoids, omega-3 LCPUFA and CLAs/CLnAs as anti-inflammatory, antiangiogenic, anti-invasive anti-cancer agents indicate common signaling pathways. Evidence in vivo and in vitro shows EPA and DHA can form endocannabinoids that: (i) are ligands for CB(½) receptors and possibly TRPV-1, (ii) have non-receptor mediated bioactivity, (iii) induce cell cycle arrest, (iii) increase autophagy and apoptosis, and (iv) augment chemotherapeutic actions in vitro. They can also form bioactive, eicosanoid-like products that appear to be non-CBR ligands but have effects on PPARs and NF-kB transcription factors. The use of cannabinoids in cancer treatment is currently limited to chemo- and radio-therapy-associated nausea and cancer-associated pain apart from one trial on brain tumours in patients. Further clinical studies are urgently required to determine the true potential of these intriguing, low toxicity compounds in cancer therapy. Particularly in view of their synergistic effects with chemotherapeutic agents similar to that observed for n-3 LCPUFA.

Dynamic simulation and visualisation of coastal erosion

A key requirement for effective coastal zone management is good knowledge and prediction of land erosion rates due to encroachment of the sea. However, in addition to demarcation of the hazard through modelling and mapping, a policy of risk mitigation necessitates significant attention should also be addressed to communicating the transient behaviour of the predictions and associated uncertainty. With climate change and sea level rise implying that historical rates of change may not be a reliable guide for the future, enhanced visualisation of the evolving coastline has the potential to improve awareness of this changing risk. This visual content is developed by linking scientific modelling with the transformation of digital elevation models, and then using GIS to integrate other spatiotemporal content. The resulting high-resolution visualisations may meet demands from decision-makers for tools to communicate scientific results more effectively, due to their realism and apparent authenticity. Nevertheless they can also produces a tension with the underlying scientific content because of the necessary extrapolation of extra detail, and the lack of established procedures to communicate the resulting uncertainty in the visualisation. Coastal managers also have concerns about releasing the visualisations to the general public. These issues are explored through analysis of future cliff erosion in Norfolk on the eastern coast of Great Britain.

Capturing coastal geomorphological change within regional integrated assessment: an outcome-driven fuzzy logic approach.

Abstract Climate change will have pervasive effects on the worlds coasts, but at broad scales these changes have typically proven difficult to analyse in a systematic manner. This paper explores an outcome-driven deductive methodology for geomorphological analysis that recognises the nonlinearity of coastal morphology and organises current knowledge and understanding using fuzzy logic concepts. Building on recent large-scale coastal investigations and with reference to a case study of the East Anglian coast, U.K., the methodology defines the active coastal system using a flexible generic classification and integrates expert opinion, using the notion of possibility, as a basis for the assessment of potential future geomorphological response to changes in sea level and sediment supply. Preliminary results for the East Anglian coast suggest that the constraining of the active coastal system by sea defences is already having, and will continue to be, a significant influence on coastal evolution irrespective of the rate of sea-level rise. Therefore, significant potential exists to guide future coastal evolution toward preferred outcomes by using this approach as a component of adaptive shoreline management. This methodology could be applied to a wide range of problems both in geomorphology and other subjects.