RESUMO
MicroRNAs are small 19-25 nucleotides which have been shown to play important roles in the regulation of gene expression in many organisms. Downregulation or accumulation of miRNAs implies either tumor suppression or oncogenic activation. In this study, differentially expressed hsa-miR-195 in hepatocellular carcinoma (HCC) was identified and analyzed. The prediction was done using a consensus approach of tools. The validation steps were done at two different levels in silico and in vitro. FGF7, GHR, PCMT1, CITED2, PEX5, PEX13, NOVA1, AXIN2, and TSPYL2 were detected with high significant (P < 0.005). These genes are involved in important pathways in cancer like MAPK signaling pathway, Jak-STAT signaling pathways, regulation of actin cytoskeleton, angiogenesis, Wnt signaling pathway, and TGF-beta signaling pathway. In vitro target validation was done for protein-L-isoaspartate (D-aspartate) O-methyltransferase (PCMT1). The co-transfection of pmirGLO-PCMT1 and pEGP-miR-195 showed highly significant results. Firefly luciferase was detected using Lumiscensor and t test analysis was done. Firefly luciferase expression was significantly decreased (P < 0.001) in comparison to the control. The low expression of firefly luciferase validates the method of target prediction that we used in this work by working on PCMT1 as a target for miR-195. Furthermore, the rest of the predicted genes are suspected to be real targets for hsa-miR-195. These target genes control almost all the hallmarks of liver cancer which can be used as therapeutic targets in cancer treatment.
Assuntos
Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patologia , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patologia , MicroRNAs/genética , Proteína D-Aspartato-L-Isoaspartato Metiltransferase/metabolismo , Regiões 3' não Traduzidas/genética , Carcinoma Hepatocelular/metabolismo , Simulação por Computador , Ontologia Genética , Humanos , Neoplasias Hepáticas/metabolismo , Luciferases/metabolismo , Células Tumorais CultivadasRESUMO
The Suez Canal region is a small fertile area in Egypt that is placed under enormous pressure of existing resources. In these areas, intensive agricultural practices are performed in combination with high population densities. The described problem is that together with agricultural practices and urbanization, Brown necked ravens (Corvus ruficollis) are in huge amounts attracted. The birds are very aggressive and showed to be a serious pest. They spread germs and diseases for human health and food production. Therefore, the birds need to be controlled. Our research focuses on identifying food preferences of raven and on the way raven control is most effective. Ravens are omnivorous birds. From our laboratory study it was seen that most preferable foods were, in descending order: fresh fishes, cow liver, crustacean, watermelons, tomatoes and yoghurt. Under field conditions where stomach content was dissected, animal matters showed to a more preferred food source than plant matters. Observations on olfactory sensitivity showed that ravens could easily locate their food. Biological observations in the field on reproduction of ravens showed that raven females lay two to six eggs. Average number of babies per nest was between one and four. Average number of raven flock before sun rise was more than 100, while it was less than 100 before sun set. The impact of mechanical, biological and chemical control was investigated. Without control, approximately flock numbers of more than 100 ravens were recorded. The average number of raven flock was 60 before mechanical control operation (nests destroyed and using net), while it was 40 after mechanical control. Results of the biological control showed that kestrel (Falco tinnunculus rupicolaeformes) predated raven babies more effective than barn owls (Tyto alba). Within the chemical control experiments, Brodifacoum (0.0005%) was most effective against ravens, followed by Zink phosphide (19%) and Methomyl (90%, carbamate compound).