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Cytotoxicity of semisynthetic derivatives of prenylflavonoids in breast carcinoma cells
Hupák, Martin ; Boušová, Iva (advisor) ; Macháček, Miloslav (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Martin Hupák Supervisor: doc. PharmDr. Iva Boušová, Ph.D. Advisor: PharmDr. Martin Ambrož, Ph.D. Title of diploma thesis: Cytotoxicity of semisynthetic derivatives of prenylflavonoids in breast carcinoma cells Cancer is a cause of a significant portion of death in the human population. In the Czech Republic, breast cancer is the most common type of cancer in women. Pharmacological treatment of cancer is often accompanied by resistance development, so new treatment options are still searched for and natural compounds are one of the sources of new structures. Prenylflavonoids present in common hop (Humulus lupulus, Cannabaceae), for example, xanthohumol, isoxanthohumol, 8-prenylnaringenin, 6- prenylnaringenin, possess a wide spectrum of biological effects. One of them is an anti- cancer effect. This diploma thesis dealt with the determination of the cytotoxic potential of semisynthetic prenylated derivatives of flavonoid naringenin in breast carcinoma cell line MCF-7. Methods used included the determination of cytotoxicity using neutral red uptake assay, analysis of apoptosis and necrosis using flow cytometry, determination of mitochondrial membrane potential using JC-1 probe, and analysis of...
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Optimization of liver spheroids preparation
Vojtěchová, Tereza ; Boušová, Iva (advisor) ; Szotáková, Barbora (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Tereza Vojtěchová Supervisor: assoc. prof. PharmDr. Iva Boušová, Ph.D. Advisor: PharmDr. Martin Ambrož, Ph.D. Title of diploma thesis: Optimization of liver spheroids preparation Various in vitro liver models play an irreplaceable role in preclinical drug evaluation. One of them is multicellular hepatic spheroids. These are 3D cell aggregates that are formed by the process of cellular aggregation. They are used in drug development and represent a promising tool for identifying new targets and drug screening for various liver diseases. The aim of this thesis was to introduce and optimize the preparation of liver spheroids by the "forced-floating" method. The liver of male and female Wistar rats was used for hepatocyte isolation and spheroid preparation. Due to the low viability of isolated hepatocytes, we tried to optimize the isolation procedure. Nevertheless, we managed to obtain only a small number of hepatocytes with sufficient viability for subsequent formation of spheroids. The spheroids were incubated with various lipogenic reagents for 7 and 14 days and their viability was monitored by measuring adenosine triphosphate (ATP) production. Incubation of spheroids with lipogenic substrates...
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Cytotoxicity of semisynthetic derivatives of prenylflavonoids in breast carcinoma cells
Hupák, Martin ; Boušová, Iva (advisor) ; Macháček, Miloslav (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Martin Hupák Supervisor: doc. PharmDr. Iva Boušová, Ph.D. Advisor: PharmDr. Martin Ambrož, Ph.D. Title of diploma thesis: Cytotoxicity of semisynthetic derivatives of prenylflavonoids in breast carcinoma cells Cancer is a cause of a significant portion of death in the human population. In the Czech Republic, breast cancer is the most common type of cancer in women. Pharmacological treatment of cancer is often accompanied by resistance development, so new treatment options are still searched for and natural compounds are one of the sources of new structures. Prenylflavonoids present in common hop (Humulus lupulus, Cannabaceae), for example, xanthohumol, isoxanthohumol, 8-prenylnaringenin, 6- prenylnaringenin, possess a wide spectrum of biological effects. One of them is an anti- cancer effect. This diploma thesis dealt with the determination of the cytotoxic potential of semisynthetic prenylated derivatives of flavonoid naringenin in breast carcinoma cell line MCF-7. Methods used included the determination of cytotoxicity using neutral red uptake assay, analysis of apoptosis and necrosis using flow cytometry, determination of mitochondrial membrane potential using JC-1 probe, and analysis of...
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Effect of flubendazole on glioblastoma multiforme in vitro and in vivo.
Šteigerová, Monika ; Boušová, Iva (advisor) ; Macháček, Miloslav (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Monika Šteigerová Supervisor: doc. PharmDr. Iva Boušová, Ph.D., RNDr. Veronika Skarková, Ph.D. Title of diploma thesis: Effect of flubendazole on glioblastoma multiforme in vitro and in vivo. Glioblastoma multiforme (GBM) is the most common malignant glioma of the central nervous system. Despite systematic complex treatment, the median survival of this malignity is 14 months. Anthelmintic and potential anticancer drug flubendazole (FLU) is recently studied for its antiproliferative effect in various tumor cells (1). The aim of our project was to investigate the effect of FLU on GBM cell lines U118MG (stabilized cell line) and GBM69 (primary cell line) in vitro, tumor growth in vivo, and its effect on mRNA and protein levels of selected markers of epithelial-mesenchymal transition and drug resistance. First-line chemotherapeutic temozolomide (TMZ) was used as a positive control. Both tested cell lines were more sensitive to FLU treatment in comparison with TMZ. Indeed, FLU caused more apparent morphological changes than TMZ in these cells. FLU significantly influenced mRNA and protein levels of E-cadherin, N-cadherin, vimentin, as well as resistance markers MGMT or MRP-1. In addition, subcutaneous...
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Epigenetic changes induced by obesity and their heredity
Roušová, Lucie ; Skálová, Lenka (advisor) ; Boušová, Iva (referee)
Obesity, a chronic disease characterized by the deposition of excessive amounts of fat in the body, is the cause of the development of many other comorbidities. In recent years, this disease appears in ever younger people. Both genetic and environmental factors are involved in the development of obesity. The link between these two factors could be epigenetic modifications. Epigenetic changes are changes in a person's phenotype without changing their genotype. These changes, which include DNA methylation and histone modification, thus affect gene expression and function, without altering the DNA sequence. The aim of my bachelor thesis was to search out which epigenetic changes are associated with obesity and what genes are affected in this way. I focused on the changes that obesity causes, but also on the changes that cause obesity. I also tried to find information about whether these genome modifications are hereditary and how many generations are possibly affected by them. From the available information, it is clear that there are a lot of epigenetic modifications that are somehow associated with obesity. For example, the changes in proopiomelanocortin, tapsin or insulin like growth factor 2 genes have been detected. Some modifications are associated with paternal obesity (e.g. the...
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Biological activity of antioxidant compounds in monocytes THP-1
Hájek, Jan ; Boušová, Iva (advisor) ; Szotáková, Barbora (referee)
Charles University in Prague University Roma Tre Faculty of Pharmacy in Hradec Králové Department of Biology Department of Biochemical Sciences Candidate: Jan Hájek Supervisor: PharmDr. Iva Boušová, Ph.D. Prof. Sandra Incerpi, Prof. Jens Z. Pedersen Title of diploma thesis: Biological activity of antioxidant compounds in monocytes THP-1 Elaboration of this work is based on the experiments carried out in the Laboratory of Physiology at University Roma Tre in Rome, under the supervision of Prof. Sandra Incerpi. The team of this laboratory studies the antioxidants of different structures and evaluates their ability to prevent the production of reactive oxygen species (ROS). These reactive species are essential for many physiological processes, but they can also have deleterious effects and participate in the development of various diseases. In this work, I dealt with a study of antioxidant properties of different polyphenolic compounds (mosloflavone, negletein, 5,6-dihydroxyflavone, baicalein, gallic acid, and compound 1625) in the cell culture of human monocytes THP-1 and also in the in vitro test. Three different methods were used: intracellular ROS determination (dichlorofluorescein assay), cytotoxicity assay, and electron paramagnetic resonance spectroscopy (EPR). Mosloflavone showed scavenging...
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Investigation of sugar modified pyrimidine nucleosides as potential inhibitors of Thymidine (TP) and Uridine (UP) phosphorylasis
Růžičková, Markéta ; Wsól, Vladimír (advisor) ; Boušová, Iva (referee)
1 Abstract of the thesis RŮŽIČKOVÁ M.; INVESTIGATION OF SUGAR MODIFIED PYRIMIDINE NUCLEOSIDES AS POTENTIAL INHIBITORS OF THYMIDINE (TP) AND URIDINE (UP) PHOSPHORYLASE Key words: uridine phosphorylase, thymidine phosphorylase, nucleoside, inhibitor, anticancer chemotherapy, substrate specifity The work deals with the investigation of novel potential inhibitors of recombinant thymidine and uridine phosphorylases from E. coli and Salmonella typhimurium. The main goals of this study are the determination of kinetic parameters for natural substrates (thymidine and uridine) for recombinant TP and UP, assessment of the role of phosphate anion in the enzymatic transformations and search for new inhibitors of TP and UP. Study of UP and TP substrate specifity and activity under the various reaction conditions is desired to obtain better view to their functions and to design novel structures of potencial inhibitors. The determination of the Michaelis constant KM and reaction rate Vmax was performed at the spectrophotometer Varian Cary-300 Bio. The representation of obtained kinetic data we used the Lineweawer-Burk plot. The next point of my work was the determination of the enzymatic activity and their substrate specifity for some pyrimidine analogues by HPLC analysis of reaction mixtures. Tested compounds...
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