3 resultados para BCL-2 FAMILY PROTEINS

em Instituto Politécnico do Porto, Portugal


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Marine cyanobacteria have been proved to be an important source of potential anticancer drugs. Although several compounds were found to be cytotoxic to cancer cells in culture, the pathways by which cells are affected are still poorly elucidated. For some compounds, cancer cell death was attributed to an implication of apoptosis through morphological apoptotic features, implication of caspases and proteins of the Bcl-2 family, and other mechanisms such as interference with microtubules dynamics, cell cycle arrest and inhibition of proteases other than caspases.

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The goal of the present paper is to analyse the classic entrepreneurship strategies (Innovation, Risk and Proactivity) in small and medium-sized businesses. However as presented in the title, the study will go further by comparing the results of those strategies in familiar and nonfamiliar businesses. This study was carried on in construction and industry sectors, in the region of Vale do Sousa, in the north of Portugal. In order to classify businesses as familiar or non-familiar types two criterion were adopted: (1) Management Control, (2) Family Employability. On the opposite to some studies that present a larger percentage of familiar businesses in national and European entrepreneurial fabric, the criterion used leaded to a larger number of non-familiar businesses (53%). The results showed that in general SMEs in this region are not following entrepreneurship strategies. Analysing the entire sample without a separation of businesses by nature (familiar/non-familiar) only proactivity showed to be more present in the managerial decisions. There is a lack of innovation and risk culture. Comparing the groups only on proactivity tests was possible to verify some differences. It was concluded that non-familiar businesses are more proactive than familiar ones. Between those groups there are no statistical differences on the means of the variables innovation and risk. At the same time some tests were conducted to test the differences on the variable entrepreneurship. The results were similar to innovation and risk strategies: There are no significant differences on entrepreneurship between these groups of businesses.

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In recent years marine biotechnology has revealed a crucial role in the future of bioindustry. Among the many marine resources, cyanobacteria have shown great potential in the production of bioactive compounds with diverse applicability. The pharmacological potential of these organisms has been one of the most explored areas in particular its antibacterial, antifungal and anticancer potential. This work was based on the assessment of potential anticancer compound E13010 F 5.4 isolated from marine cyanobacteria strain Synechocystis salina LEGE 06099. Thus the aim of this work was to explore molecular and biochemical mechanisms underlying the bioactivity detected in human cancer cells, specifically in lines RKO colon carcinoma and HT-29. The isolation of the compound was performed from biomass obtained by large-scale culture. To obtain the compound fractionation was carried and confirmation and isolation performed by Nuclear Magnetic Resonance (NMR), Thin Layer Chromatography (TLC) and High-Performance Liquid Chromatography (HPLC). Cell viability assays were performed based on reduction of 3- (4,5-dimetiltiaziol-2-yl) -2,5-diphenyltetrazolium bromide (MTT) to assess the cytotoxic potential of the compound. From the battery of cell lines RKO (colon carcinoma), HT-29 (colorectal adenocarcinoma), MG-63 (osteosarcoma) and T47D (breast carcinoma) the cell lines RKO and HT-29 were selected for elucidation of mechanisms of cytotoxicity. For the elucidation of the mechanisms involved in cytotoxicity the cell lines RKO and HT29 were exposed to the compound. A genomic approach based in the mRNA expression of genes involved in apoptosis and cell cycle by Real-Time PCR and a proteomic approach based on the separation of proteins by two-dimensional electrophoresis (2DGE) was performed. For mRNA expression were selected the genes RPL8, HPRT1, VDAC, SHMT2, CCNE, CCNB1, P21CIP, BCL-2 and BAD and for proteomics isoelectric focussing between 3 – 10 and molecular weight of 19 – 117 kDa separated by polyacrylamide gels (2DGE). The MTT results confirmed the reduction of the cell viability. The RT-PCR results for the expression of genes studied were not yet fully elucidative. For the cell line RKO there was a significant reduction in the expression of the gene P21CIP, and a tendency for reduction in the BAD gene expression and for increased expression of gene CCNB1, pointing to an effort for cell proliferation. In HT-29 cell line, there was a tendency for increase in the expression of P21CIP and BAD, which may explain the reduction in cell viability. The 2DGE results indicate proteomic patterns with differentially altered spots in the treated and control cells with both qualitative and quantitative differences, and differences in response between the RKO and HT-29 cell lines.