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The group of DHRS enzymes and their role in human organism
Zamazalová, Alžběta ; Zemanová, Lucie (advisor) ; Štambergová, Hana (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Alžběta Zamazalová Supervisor: RNDr. Lucie Zemanová, Ph.D. Title of diploma thesis: The group of DHRS enzymes and their role in human organism Human enzymes from dehydrogenase/reductase SDR family (DHRS) include 17 members that belong to the large short-chain dehydrogenase/reductase superfamily. The short-chain dehydrogenase/reductase superfamily contains 80 human members, some of them are important and well described enzymes such as 11β-hydroxysteroid dehydrogenase, carbonyl reductase 1 and 17β-hydroxysteroid dehydrogenase. However there are many poorly described or undescribed members of this superfamily, including group DHRS. There is just a little information about proteins DHRS1, DHRS12, DHRS13, DHRS4L1, DHRS4L2. Proteins DHRS2, DHRS4, DHRS6, DHRS7, DHRS7B, DHRS7C, DHRS8, DHRS9, DHRS10, DHRS11 and DHRSX are better described because at least subcellular localization and substrate (except DHRSX) is known, and therefore an initial opinion on their possible function can be formed. The DHRS3 is the only one enzyme of group DHRS that has been tested in vivo. DHRS3 could play a significant role in fetal development. For DHRS3, but also for the other DHRS enzymes, first knowledge of...
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Production of the ovarian hormones in breast tumours.
Štambergová, Hana ; Hochmann, Jiří (advisor) ; Dršata, Jaroslav (referee)
1 SUMMARY The breast cancer affects over one million women all over the world every year. Approximately 60 - 80 % of these tumours are declared as hormone sensitive, which means they are able to respond on a stimulation of estrogens that work as cocancerogens. The breast cancer incidence increases according to an age, a majority of new cases is detected during the postmenopausal period when the ovaries ceased to be functional. In postmenopausal patients the local estrogen production becomes a considerable stimulating growth factor of tumours. In this thesis we have brought together a review of enzymes participating on the intratumoral biosynthesis of estrogens or estradiol. The enzymes responsible for the local synthesis include predominantly the aromatase, 17β-hydroxysteroid dehydrogenases and steroid sulphatase. We have specified by each type of the enzymes its biochemistry, biological characteristics and also its engagement in a breast cancerogenesis as well as their advantage as a pharmacological target. We have also placed pieces of information about members of the AKR1C subfamily. Professor Wsól from the department of biochemistry sciences of our faculty focuses on their study (a participation on a biodegradation of carbonyl groups xenobiotics). AKR1C3 (also called 17β-hydroxysteroid dehydrogenase...
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The group of DHRS enzymes and their role in human organism
Zamazalová, Alžběta ; Zemanová, Lucie (advisor) ; Štambergová, Hana (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Alžběta Zamazalová Supervisor: RNDr. Lucie Zemanová, Ph.D. Title of diploma thesis: The group of DHRS enzymes and their role in human organism Human enzymes from dehydrogenase/reductase SDR family (DHRS) include 17 members that belong to the large short-chain dehydrogenase/reductase superfamily. The short-chain dehydrogenase/reductase superfamily contains 80 human members, some of them are important and well described enzymes such as 11β-hydroxysteroid dehydrogenase, carbonyl reductase 1 and 17β-hydroxysteroid dehydrogenase. However there are many poorly described or undescribed members of this superfamily, including group DHRS. There is just a little information about proteins DHRS1, DHRS12, DHRS13, DHRS4L1, DHRS4L2. Proteins DHRS2, DHRS4, DHRS6, DHRS7, DHRS7B, DHRS7C, DHRS8, DHRS9, DHRS10, DHRS11 and DHRSX are better described because at least subcellular localization and substrate (except DHRSX) is known, and therefore an initial opinion on their possible function can be formed. The DHRS3 is the only one enzyme of group DHRS that has been tested in vivo. DHRS3 could play a significant role in fetal development. For DHRS3, but also for the other DHRS enzymes, first knowledge of...
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Determination of the expression of enzymes DHRS7B and DHR7C in human tissues
Kozáková, Klára ; Wsól, Vladimír (advisor) ; Štambergová, Hana (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Klára Kozáková Supervisor: prof. Ing. Vladimír Wsól, Ph.D. Title of diploma thesis: Determination of the expression of enzymes DHRS7B and DHRS7C in human tissues Dehydrogenase/reductase (SDR family) member 7B (DHRS7B) and 7C (DHRS7C) are human enzymes that belong to the short chain dehydrogenase/reductase (SDR) superfamily. This very old and extensive superfamily includes members, which play an important role in different physiological and pathological processes. In spite of that the SDR enzymes are currently more studied, still about half of these enzymes are only poorly characterized, such as DHRS7B and DHRS7C. The aim of this study was the determination of the expression profiles of human enzymes DHRS7B a DHRS7C at mRNA and protein level and thus brings new knowledge of these enzymes. Expression of enzymes was tested in human tissues. RNA was isolated from the samples and used for transcription to cDNA. Obtained cDNA was utilized as a template for qPCR with fluorescence SYBR GREEN I detection. Determination of enzymes at protein level was done in homogenates of human tissues by using a Western blotting with chemiluminescence detection. The level of expression mRNA DHRS7B was strongest in testes...
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Impact of selected sesquiterpenes on biotransformation and antioxidant enzymes in hepatocytes and cancer cell line MCF-7
Auterská, Lenka ; Svobodová, Hana (advisor) ; Štambergová, Hana (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Lenka Auterská Supervisor: PharmDr. Hana Bártíková, Ph.D Title of diploma thesis: Impact of selected sesquiterpenes on biotransformation and antioxidant enzymes in hepatocytes and cancer cell line MCF-7 A breast cancer belongs among the most common cancer diseases of women and its incidence keeps increasing. The highest percentage is represented by a ductal carcinoma, which originates in the cells of the milk ducts and from there it can spread to other tissues. Nowadays, there are several cell lines, which serve for research purposes in the field of cancer therapy as a model for in vivo and in vitro studies. Particularly the MCF-7 cell line has been proved as a significant source of information about the response of cancer tissues to drug administration. It has been proved that content components of essential oils are able to inhibit cancer cell proliferation, induce apoptosis, suppress abiogenesis and metastases, and enhance the effect of chemotherapy. Besides other things, numerous studies have confirmed their effect on the activity of enzymes involved in a body protection against free radicals and oxidative stress. The aim of this thesis was to determine the cytotoxic effect of...
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Molecular cloning of YinP gene from Leishmania major using two red fluorescent pXG-mCherry plasmids. Valuable tools for gene expression location.
Musilová, Kateřina ; Wsól, Vladimír (advisor) ; Štambergová, Hana (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Kateřina Musilová Supervisor: Prof. Ing. Vladimír Wsól, Ph.D. Tutor: Prof. Paul Nguewa, Ph.D. Title of diploma thesis: Molecular cloning of YinP gene from Leishmania major using two red fluorescent pXG-mCherry plasmids. Valuable tools for gene expression location. In the 21st century, leishmaniasis remains a major health problem in numerous de- veloping countries. Around 2 million cases of leishmaniasis are reported every year and estimated mortality is over 20,000 deaths annually. Antileishmanial drugs are often unaf- fordable for affected people and display severe toxic side effects. Potent human vaccines are not available. This, together with increasing resistance, is a reason why new effective, safe, and affordable medicines are greatly needed. Leishmaniasis is caused by Leishmania species. These parasites are transmitted by phlebotomine sand flies, which also provide to the leishmania environment necessary for their development into infective forms. The process of transformation into a stage in- fective for vertebrate hosts is called metacyclogenesis. Nowadays, genes, enzymes, and proteins possibly exhibiting a function in the metacyclogenesis are extensively examined. One of the...
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Optimization of transfection of eukaryotic cell lines
Šplíchalová, Iva ; Zemanová, Lucie (advisor) ; Štambergová, Hana (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Iva Šplíchalová Supervisor: RNDr. Lucie Škarydová, Ph.D. External supervisor: Mgr. Vojtěch Vejvoda, Ph.D. Title of Master thesis: Optimization of Transfection of Eukaryotic Cells An optimization of a transfection is a process of searching for the best conditions for the transfection. Expression systems corresponding to concrete requirements are chosen first of all. A speed, an economy and a functionality of prepared recombinant protein that can be influenced by posttranslational modifications must be considered. Expression vectors are used to insert genes of the interest into expression systems. The goal of this diploma thesis was to optimize conditions for transfection of eukaryotic cell lines CHO PF a HEK293 using of three commercially available kits. It was followed by a comparison of these transfection efficiency with standard procedures that are used in the company Generi Botech s.r.o. A plasmid pMaxGFP was used for the optimization. A cell counting in Neubaurer's chamber and a quantification of GFP in cell lysate were used to evaluate the efficiency of the transfection. Commercially available kits ...
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Inhibition study of human 17β-HSD3
Poláková, Lenka ; Štambergová, Hana (advisor) ; Zemanová, Lucie (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Lenka Poláková Supervisor: PharmDr. Hana Štambergová, Ph.D. Title of diploma thesis: Inhibition study of human 17β-HSD3 Enzyme 17β-hydroxysteroid dehydrogenase 3 belonging to the short-chain dehydrogenase/reductase superfamily is involved mainly in reduction of 4-androstane-3,17- dione to testosterone. Any changes in amount of testosterone can result in illness, such as the male pseudhermaphroditism or the prostate cancer. For this reason studies of inhibion effects of various substances have been doing as they could be used in the therapy such as illnesses. Identification of the pharmacologically interesting substances was performed in vitro through the model system. Cell line Sf9 was used to prepare a recombinant form of the target enzyme. Reduction activity of this enzyme was verified towards androstene-4,17-dione. Then screening of potential inhibitors, natural substances: flavonoids and alkaloids, was performed. IC50 value was defined at substances with 100% inhibition effect (2-hydroxyflavanone, biochanine A, fumariline).
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