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Proteomic analysis of posttranslation modifications in breast cancer cell line profiles
Predná, Nikola ; Laštovičková,, Markéta (oponent) ; Strouhalová,, Dana (vedoucí práce)
Estrogen and progesterone receptors, as well as HER2 protein, are currently the most clinically useful metabolic markers in breast cancer. These markers allow for the determination of the type of tumor and its best treatment options. However, one of the most aggressive types of this disease, triple-negative breast cancer (TNBC), lacks these clinically established biomarkers. This means that hormone therapy or targeted drugs are not an option, leaving fewer treatment options to choose from. In order for new tailored drugs to be developed, the understanding of the molecular basis of the disease is crucial. Recently, many studies aim their search for biomarkers at the protein level using proteomics. Proteins, notably their post-translational modifications (PTM), are at the core of many cellular events and their uncovering may help in the understanding of breast cancer mechanisms.In order to discover the molecular features of TNBC, this study aims to compare proteomic data of untreated cancer cell lines with cells that underwent retinoid therapy. The focus will be on the PTMs, notably glycosylation and phosphorylation, of Vimentin and CD44, which were proposed as potential TNBC biomarkers in previous studies. Protein separation will be carried out using 1D and 2D gel electrophoresis or by SEC-HPLC. The samples will also be subdued to enzymatic cleavage before being identified using MALDI-TOF Mass Spectrometry. In the case of phosphoprotein selective capture, enrichment will be performed by affinity chromatography using TiO2 phosphopeptide enrichment tips (TopTip). Glycosylated proteins will be enriched using WGA lectin affinity based chromatography. Proteins with significant differences in PTMs between the treated and untreated cells will be evaluated using protein databases (MASCOT, STRING, and more). The data acquired from the study will eventually be used to propose potential biomarkers for TNBC.
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Proteomic approach for the study of cancer cell line profiles.
Predná, Nikola ; Langová, Denisa (oponent) ; Strouhalová,, Dana (vedoucí práce)
Triple-negative breast cancer (TNBC), known to be an aggressive subtype of breast cancer, has a dismal prognosis and limited treatment options. As of now, chemotherapy is considered the main treatment option. In order for new effective drugs to be developed, it is crucial to understand the molecular basis of triple-negative breast cancer. As a result, a lot of research on potentially active agents for this particular type of breast cancer have been conducted. Recently, many studies on the matter are carried out using proteomics; a means of studying the proteomes of cancer cells. Cancer cells contain key differences in proteins that regulate the mechanisms of the cell. Eventually, mapping these mechanisms can allow to define the state of an organism. This thesis focuses on the proteomic study of TNBC cells and compares untreated cells with cells that have underwent retinoid therapy. Protein and peptide separation was successfully performed using 1D and 2D gel electrophoresis. In addition, the samples were subjected to enzymatic cleavage of selected proteins which were then identified using MALDI-TOF Mass Spectrometry (MS). Proteins playing a role in the process of epithelial–mesenchymal transition (EMT) were then quantified and compared between the samples.
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Proteomic analysis of posttranslation modifications in breast cancer cell line profiles
Predná, Nikola ; Laštovičková,, Markéta (oponent) ; Strouhalová,, Dana (vedoucí práce)
Estrogen and progesterone receptors, as well as HER2 protein, are currently the most clinically useful metabolic markers in breast cancer. These markers allow for the determination of the type of tumor and its best treatment options. However, one of the most aggressive types of this disease, triple-negative breast cancer (TNBC), lacks these clinically established biomarkers. This means that hormone therapy or targeted drugs are not an option, leaving fewer treatment options to choose from. In order for new tailored drugs to be developed, the understanding of the molecular basis of the disease is crucial. Recently, many studies aim their search for biomarkers at the protein level using proteomics. Proteins, notably their post-translational modifications (PTM), are at the core of many cellular events and their uncovering may help in the understanding of breast cancer mechanisms.In order to discover the molecular features of TNBC, this study aims to compare proteomic data of untreated cancer cell lines with cells that underwent retinoid therapy. The focus will be on the PTMs, notably glycosylation and phosphorylation, of Vimentin and CD44, which were proposed as potential TNBC biomarkers in previous studies. Protein separation will be carried out using 1D and 2D gel electrophoresis or by SEC-HPLC. The samples will also be subdued to enzymatic cleavage before being identified using MALDI-TOF Mass Spectrometry. In the case of phosphoprotein selective capture, enrichment will be performed by affinity chromatography using TiO2 phosphopeptide enrichment tips (TopTip). Glycosylated proteins will be enriched using WGA lectin affinity based chromatography. Proteins with significant differences in PTMs between the treated and untreated cells will be evaluated using protein databases (MASCOT, STRING, and more). The data acquired from the study will eventually be used to propose potential biomarkers for TNBC.
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Proteomic approach for the study of cancer cell line profiles.
Predná, Nikola ; Langová, Denisa (oponent) ; Strouhalová,, Dana (vedoucí práce)
Triple-negative breast cancer (TNBC), known to be an aggressive subtype of breast cancer, has a dismal prognosis and limited treatment options. As of now, chemotherapy is considered the main treatment option. In order for new effective drugs to be developed, it is crucial to understand the molecular basis of triple-negative breast cancer. As a result, a lot of research on potentially active agents for this particular type of breast cancer have been conducted. Recently, many studies on the matter are carried out using proteomics; a means of studying the proteomes of cancer cells. Cancer cells contain key differences in proteins that regulate the mechanisms of the cell. Eventually, mapping these mechanisms can allow to define the state of an organism. This thesis focuses on the proteomic study of TNBC cells and compares untreated cells with cells that have underwent retinoid therapy. Protein and peptide separation was successfully performed using 1D and 2D gel electrophoresis. In addition, the samples were subjected to enzymatic cleavage of selected proteins which were then identified using MALDI-TOF Mass Spectrometry (MS). Proteins playing a role in the process of epithelial–mesenchymal transition (EMT) were then quantified and compared between the samples.
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Topická a systémová léčba acne vulgaris
Ackermannová, Veronika ; Hrdina, Radomír (vedoucí práce) ; Vopršalová, Marie (oponent)
Univerzita Karlova Farmaceutická fakulta v Hradci Králové Katedra farmakologie a toxikologie Studentka: Veronika Ackermannová Školitel: prof. MUDr. Radomír Hrdina, CSc. Název diplomové práce: Topická a systémová léčba acne vulgaris Acne vulgaris je kožní onemocnění postihující vlasové folikuly a mazové žlázy. Projevuje se zvýšenou tvorbou mazu, nezánětlivými (komedony) a zánětlivými lézemi (papuly, pustuly, noduly, cysty). Vyskytuje se převážně u adolescentů, nicméně může přetrvávat i do dospělosti. Jedná se o multifaktoriální onemocnění, na jehož vzniku má podíl více faktorů (vnitřní a vnější podněty). Mezi hlavní patogenetické faktory patří zvýšená tvorba kožního mazu, hyperkeratóza, osídlení bakteriemi P. acnes a přítomný zánět. Pro zvolení správné a účinné terapie je nutné nejprve diagnostikovat daný typ akné, jelikož neexistuje pouze jeden typ. Typů a variant akné je veliké množství, a ačkoliv se projevují podobnými projevy (zásah do folikulů mazových žláz), jejich příčina vzniku se často liší. Neexistuje jednotný systém klasifikace acne vulgaris a mezi autory se liší. Někteří dělí acne vulgaris podle závažnosti na mírnou, středně závažnou a závažnou formou, jiní na komedonickou, papulopustulózní, nodulocystickou a konglobátní akné. Léčivé přípravky užívané v terapii akné obsahují látky z...
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Využití proteomiky pro monitorování potenciálních biomarkerů rakoviny prsu
Garcia N Dua, Felicita ; Ing.Anna Kubesová, Ph.D. (oponent) ; Flodrová, Dana (vedoucí práce)
Nádor prsu je jedním z nejčastějších maligních karcinomů u žen a prognóza úzce souvisí s tím, v jakém stádiu kancerogeneze je proces zachycen. V posledních letech je pozornost soustřeďována na hledání nových látek, jež by snížily negativní dopady, které s sebou nese chemoterapie. Bylo zjištěno, že jednou z efektivních látek v boji s karcinomem jsou retinoidy, které mají schopnost inhibovat nádory prsní žlázy v časných stádiích nemoci. Tato práce se zabývá proteomickými studiemi buněk karcinomu prsu a porovnáváním obsahu bílkovin obsažených v karcinomových buňkách, které byly podrobeny retinoidové léčbě. Úspěšně bylo využito několika separačních metod jako 1D a 2D elektroforéza, chromatografie na reverzní fázi a gelová permeační chromatografie. K tomu byly vzorky ještě podrobeny enzymatickému štěpení vybraných proteinů a jejich identifikaci pomocí MALDI-TOF MS. 2D gelová elektroforéza odhalila výskyt skupiny proteinů řadících se mezi molekulární chaperony, které jsou dnes považovány za významné specifické biomarkery nejen karcinomu prsu.
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