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Functional analysis of differentially expressed miRNAs in HPV positive and HPV negative cell lines derived various anatomical localization
Krbušek, David ; Saláková, Martina (advisor) ; Dostálová Merkerová, Michaela (referee)
MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression. MiRNA expression profiles showed several differentially expressed miRNAs specific to human papillomavirus (HPV)-associated tumors. These miRNAs include miR-139-5p, which has reduced expression in these tumors. In this thesis, the role of miR-139-5p was studied in vitro on cell lines that were HPV positive (CRL-3240, SiHa) and HPV negative (FaDu, C-33A). Cell lines were transfected with mimic miRNA, the ability of cells to proliferate and migrate was then studied. Cell proliferation was studied using MTT assay, while Scratch wound healing assay and transwell assay were used to evaluate the migratory abilities of the cells. Mediator RNAs (mRNAs) of target genes of miR-139-5p were predicted using TargetScan and miRDB databases. The change in gene expression of target mRNAs, as well as the verification of the successful increase in miRNA expression in the cell lines, was verified using RT-qPCR. Increase of miR-139-5p expression in all used cell lines did not lead to statistically significant changes (p≤0.05) in proliferative or migratory abilities. The mRNAs of FOS, JUN, KIF13A and GDF10 genes were selected as targets of miR-139-5p. Transfected cell lines did not show a noticable reduction in the expression of the target mRNAs....
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The role of microRNA in physiology and pathology
Janáková, Barbora ; Matoušková, Petra (advisor) ; Pávek, Petr (referee) ; Dostálová Merkerová, Michaela (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Mgr. Barbora Hanousková Supervisor: doc. Ing. Petra Matoušková, Ph.D. Title of Doctoral Thesis: The role of microRNA in physiology and pathology MicroRNAs (miRNAs) are small single-stranded non-coding RNA molecules that play an important role in the regulation of gene expression. They are evolutionarily highly conserved and are present in the genome of all eukaryotic organisms, suggesting their importance in physiological processes. Due to the tissue specificity and their involvement in the pathogenesis of various diseases, miRNAs have been widely studied in connection with their potential use as specific and early biomarkers, or as a therapeutic target. The doctoral thesis, written in the form of an annotated set of publications, dealt with the study of miRNAs in various systems in vitro and in vivo, focusing on their role in adipogenesis and their use as biomarkers of pathological conditions. The introduction summarizes the theoretical information on epigenetic regulation with a more detailed focus on miRNAs, followed by comments on the author's individual publications and brief conclusions. In two studies using mouse models, several miRNAs were identified to have their expression profile...
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The role of microRNA in physiology and pathology
Janáková, Barbora ; Matoušková, Petra (advisor) ; Pávek, Petr (referee) ; Dostálová Merkerová, Michaela (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Mgr. Barbora Hanousková Supervisor: doc. Ing. Petra Matoušková, Ph.D. Title of Doctoral Thesis: The role of microRNA in physiology and pathology MicroRNAs (miRNAs) are small single-stranded non-coding RNA molecules that play an important role in the regulation of gene expression. They are evolutionarily highly conserved and are present in the genome of all eukaryotic organisms, suggesting their importance in physiological processes. Due to the tissue specificity and their involvement in the pathogenesis of various diseases, miRNAs have been widely studied in connection with their potential use as specific and early biomarkers, or as a therapeutic target. The doctoral thesis, written in the form of an annotated set of publications, dealt with the study of miRNAs in various systems in vitro and in vivo, focusing on their role in adipogenesis and their use as biomarkers of pathological conditions. The introduction summarizes the theoretical information on epigenetic regulation with a more detailed focus on miRNAs, followed by comments on the author's individual publications and brief conclusions. In two studies using mouse models, several miRNAs were identified to have their expression profile...
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Small noncoding RNAs and their prognostic value in myelodysplastic syndromes
Hruštincová, Andrea ; Dostálová Merkerová, Michaela (advisor) ; Stopka, Tomáš (referee) ; Vymetálková, Veronika (referee)
Myelodysplastic syndromes (MDS) are a heterogeneous group of bone marrow (BM) disorders characterized by ineffective haematopoiesis, BM dysplasia, and peripheral blood cytopenia. In recent years, substantial progress has been made towards understanding the molecular pathogenesis of MDS that has brought new possibilities in MDS diagnostics, prognostics, and treatment. Small noncoding RNAs (sncRNAs), especially microRNAs (miRNAs), are in the field of scientific interest in terms of their expression, function, role in disease development, and potential utilization as disease biomarkers. Special attention has been focused on extracellular sncRNAs present in blood circulation, so called 'circulating' sncRNAs, which may become easily accessible biomarkers of disease state or risk of progression. We have conducted several studies on intracellular and extracellular sncRNA profiles of CD34+ BM cells and blood plasma, respectively, from MDS patients using microarrays or next generation sequencing (NGS). We aimed to identify specific sncRNA profiles associated with MDS and search for sncRNA biomarkers predictive of the patient prognosis and response to treatment with azacitidine (AZA). Another goal was to characterize and compare circulating sncRNA profiles of two different extracellular materials, total...
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Circadian activity of tadpoles of Phyllobates vittatus.
DOSTÁLOVÁ, Michaela
Tadpoles are known to behaviorally respond to visual, auditory or chemical cues. I examined the influence of group size and light conditions on the activity of Phyllobates vittatus. I performed laboratory experiments to measure locomotor activity. Activity levels of tadpoles were higher in the groups. Light conditions did not influence activity level of tadpoles. My results suggest that the group size is one of the main factors influencing tadpole locomotor activity of Phyllobates vittatus.
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