Transcriptional down-regulation of the retinoblastoma protein is associated with differentiation and apoptosis in human colorectal epithelial cells

The aim of this study was to investigate the regulation of Rb protein expression in relation to increased differentiation and induction of apoptosis in colonic epithelial cells. In vivo, Rb protein expression was found to be down-regulated towards the top of the normal colonic crypt, coincident with the region of differentiation and apoptosis, but highly expressed in colonic carcinoma tissue. Using in vitro models to study the regulation of Rb expression in pre-malignant colonic epithelial cells, we have been able to show for the first time that Rb protein expression is transcriptionally down-regulated in differentiated pre-malignant cells (in post-confluent cultures) but not in malignant colorectal epithelial cells. Furthermore, suppression of rb protein function by the HPV-E7 viral oncoprotein increased both spontaneous and DNA damage-induced apoptosis. These results suggest that Rb is able to act as a survival factor in colonic epithelial cells by suppressing apoptosis, and that over-expression of pRb in colorectal tumour cells can cause a loss of sensitivity to apoptotic signalling, resulting in aberrant cell survival and resistance to therapy. © 2001 Cancer Research Campaign http://www.bjcancer.co

The actin-bundling protein fascin is overexpressed in colorectal adenomas and promotes motility in adenoma cells in vitro

Background: Fascin is overexpressed in many cancers, including colorectal, but its role in the malignant transformation of benign colorectal adenomas is unclear. Methods: Immunohistochemical analysis of fascin expression was carried out in resected human colorectal adenoma specimens. The effects of forced overexpression of fascin on adenoma cell motility were also analysed. Results: We show fascin overexpression in adenomas increasing with tumour size, histological type, and degree of dysplasia and increased cell motility in adenoma cell lines following fascin transfection. Conclusion: These data suggest an important role for fascin in the malignant progression of colorectal tumours

β-catenin negatively regulates expression of the prostaglandin transporter PGT in the normal intestinal epithelium and colorectal tumour cells: A role in the chemopreventive efficacy of aspirin

Background: Levels of the pro-tumorigenic prostaglandin PGE 2 are increased in colorectal cancer, previously attributed to increased synthesis through COX-2 upregulation and, more recently, to decreased catabolism. The functionally linked genes 15-prostaglandin dehydrogenase (15-PGDH) and the prostaglandin transporter PGT co-operate in prostaglandin degradation and are downregulated in colorectal cancer. We previously reported repression of 15-PGDH expression by the Wnt/β-catenin pathway, commonly deregulated during early colorectal neoplasia. Here we asked whether β-catenin also regulates PGT expression. Methods: The effect of β-catenin deletion in vivo was addressed by PGT immunostaining of β-catenin/lox-villin-cre-ERT2 mouse tissue. The effect of siRNA-mediated β-catenin knockdown and dnTCF4 induction in vitro was addressed by semi-quantitative and quantitative real-time RT-PCR and immunoblotting. Results: This study shows for the first time that deletion of β-catenin in murine intestinal epithelium in vivo upregulates PGT protein, especially in the crypt epithelium. Furthermore, β-catenin knockdown in vitro increases PGT expression in both colorectal adenoma-and carcinoma-derived cell lines, as does dnTCF4 induction in LS174T cells.Conclusions:These data suggest that β-catenin employs a two-pronged approach to inhibiting prostaglandin turnover during colorectal neoplasia by repressing PGT expression in addition to 15-PGDH. Furthermore, our data highlight a potential mechanism that may contribute to the non-selective NSAID aspirins chemopreventive efficacy. © 2012 Cancer Research UK All rights reserved

Giant liposarcoma of the back with 4 types of histopathology: a case report

The incidence of soft tissue tumours, both malignant and benign, is very common. However, the coexistence of 4 types of histopathology is rare and the aim of this article is to present one treated in our Department. An 87-year-old Greek man was treated in our Department for a huge tumour on his back, under local anaesthesia. The pathology report of the specimen referred 4 types of neoplasia. This case represents this incidence in a giant liposarcoma of the back

Nutrient stress alters the glycosylation status of LGR5 resulting in reduced protein stability and membrane localisation in colorectal tumour cells: implications for targeting cancer stem cells

BACKGROUND LGR5 is an important marker of intestinal stem cells and performs its vital functions at the cell membrane. Despite the importance of LGR5 to both normal and cancer stem cell biology, it is not known how microenvironmental stress affects the expression and subcellular distribution of the protein. METHODS Nutrient stress was induced through glucose starvation. Glycosylation status was assessed using endoglycosidase or tunicamycin treatment. Flow cytometry and confocal microscopy were used to assess subcellular distribution of LGR5. RESULTS Glucose deprivation altered the glycosylation status of LGR5 resulting in reduced protein stability and cell surface expression. Furthermore, inhibiting LGR5 glycosylation resulted in depleted surface expression and reduced localisation in the cis-Golgi network. CONCLUSIONS Nutrient stress within a tumour microenvironment has the capacity to alter LGR5 protein stability and membrane localisation through modulation of LGR5 glycosylation status. As LGR5 surface localisation is required for enhanced Wnt signalling, this is the first report to show a mechanism by which the microenvironment could affect LGR5 function

Problems associated with direct displacement-based design of concrete bridges with single-column piers, and some suggested improvements

Currently available displacement-based design (DBD) procedures for bridges are critically evaluated with a view to identifying extensions and/or modifications of the procedure, for it to be applicable to final design of a fairly broad class of bridges. An improved direct DBD procedure is presented, including a suite of comprehensive design criteria and proper consideration of the degree of fixity of the pier top. The design of an overpass bridge (originally designed to a current European Code), applying the improved ‘direct’ displacement-based design (DDBD) procedure is presented and both ‘conventional’ and displacement-based designs are assessed using non-linear response-history analysis (NLRHA); comparisons are made in terms of both economy and seismic performance of the different designs. It is seen that DDBD provided a more rational base shear distribution among piers and abutments when compared to the force-based design procedure and adequately captured the displacement pattern, closely matching the results of the more rigorous NLRHA

Multimedia e-learning on technologies for efficient water use

One of the Millennium Development Goals is “To ensure environmental sustainability”, with a special task dedicated to halve by 2015 the proportion of people without sustainable access to safe drinking water. This task collides with alarming studies that, for instance, foresee an acute water shortage in India and other Asian countries by 2050. In this conflicting context we present ED-WAVE, a developing project funded by the Asia-Link Programme, participating six countries (India, Sri Lanka, Thailand, Greece, Spain and Finland). Its goal is to create a sustainable international cooperation framework and to develop a multimedia tool for e-Learning on technologies for conservation, reclamation and reuse of natural resources. We therefore use IT tools aimed at improving people’s education, including analysis of current water use status in Asia and Europe, review of potential technological interventions, simulation of impacts of specific technology interventions and translation of knowledge into electronic teaching material

Performance-Based Seismic Design and Assessment of Bridges

Current trends in the seismic design and assessment of bridges are discussed, with emphasis on two procedures that merit some particular attention, displacement-based procedures and deformation-based procedures. The available performance-based methods for bridges are critically reviewed and a number of critical issues are identified, which arise in all procedures. Then two recently proposed methods are presented in some detail, one based on the direct displacement-based design approach, using equivalent elastic analysis and properly reduced displacement spectra, and one based on the deformation-based approach, which involves a type of partially inelastic response-history analysis for a set of ground motions and wherein pier ductility is included as a design parameter, along with displacement criteria. The current trends in seismic assessment of bridges are then summarised and the more rigorous assessment procedure, i.e. nonlinear dynamic response-history analysis, is used to assess the performance of bridges designed to the previously described procedures. Finally some comments are offered on the feasibility of including such methods in the new generation of bridge codes