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The study of the anticancer effect of platinum complexes PHEN-BEQU
Večeřová, Michaela ; Prachařová,, Jitka (referee) ; Kostrhunová,, Hana (advisor)
Cancer represents a significant threat to society, and although there is a wide range of solutions to this problem available today, none are without risks. The task of the bachelor’s thesis was to study the anti-tumor effect of newly synthesized platinum complexes of the PHEN-BEQU group. First, an MTT test was performed, and the IC50 values of individual complexes were obtained as a result. Cytotoxicity was also measured using the CellTiter-Glo® assay on spheroids - 3D cellular models capable of better simulating the environment of a real tumor. Furthermore, the accumulation of complexes in cells and nuclei was measured, and the type of cell death was determined. The results were correlated with the content of lipophilic groups, and their effects were evaluated.
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Analysis of Microscopic Images of Cancer Cells
Vičar, Tomáš ; Matula,, Petr (referee) ; Sladoje, Natasa (referee) ; Kolář, Radim (advisor)
Tato disertační práce je zaměřena na analýzu různých forem mikroskopických obrazových dat nádorových buněk (statické 2D snímky, statické 3D obrazy, 2D časosběrné zobrazování živých buněk). Hlavní pozornost je věnována datům získaným koherencí řízeným holografickým mikroskopem, který je relativně novou modalitou schopnou kotrastních záznamů živých buněk bez barvení (label-free) a poskytuje kvantitativní informaci (kvantitativní fázové zobrazení - QPI). V práci je popsán základní postup analýzy těchto snímků a jsou vytvářeny nové metody a zdokonalovány metody pro jednotlivé kroky této analýzy. Největší část práce je věnována segmentaci buněk, kde jsou shrnuty klasické metody i metody založené na hlubokém učení. Jsou také vyvinuty nové metody vhodné právě pro QPI data. Část práce je také věnována segmentaci 3D fluorescenční jader a detekci DNA zlomů pomocí hlubokého učení. Práce se zabývá i dalším zpracování v podobě sledování buněk, extrakce příznaků a následné analýze, kde je detekována buněčná smrt a jsou vytvořeny vhodné interpretovatelné příznaky pro klasifikaci buněčné smrti na apoptickou a lytickou. Celkově tato práce přispívá k rozvoji jednotlivých kroků analýzy obrazu nádorových buněk a odráží současný pokrok v oblasti analýzy obrazu, zejména přístupy hlubokého učení, což je také demonstrováno na několika výzkumných aplikacích.
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Analysis of changes in the phenotype of tumour cells induced by migrastatics in quantitative phase imaging
Kolínková, Veronika ; Netíková,, Irena Štenglová (referee) ; Veselý, Pavel (advisor)
This thesis focuses on the observation of living cells using the non-invasive methods of quantitative phase imaging (hiQPI). The imaging is enabled by a coherence-controlled holographic microscope (CCHM) developed in the Laboratory of Experimental Biophotonics at the VUT. Using this imaging technology, morphological changes of A549 and MCF7 after application of potential migrastatic drugs tumor cells are evaluated in the experimental part of the thesis. Migrastatics are defined as already approved drugs that could prevent the migration of cancer cells from the primary tumor and thus prevent metastasis. The RAC-GM (Rapid Assessment of Cell Growth and Migration in Vitro) method was chosen to assess their effect on tumor cells.
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Therapy of difficult-to-treat pancreatic tumours with new metallopharmaceuticals and their mechanism of action
Švitelová, Marie ; Prachařová,, Jitka (referee) ; PhD, Vojtěch Novohradský, (advisor)
The issue of tumor diseases and anticancer therapy is extremely current nowadays, in which is all acquired knowledge a great benefit to society. Despite the extraordinary development of this field, the available treatment approaches for cancer are still very limited by a number of factors, that motivate research teams to find targeted, effective therapy. The main aim of this bachelor work was to test an in vitro cytotoxicity of newly synthesized enantiomers [Pt(OXA)(1,2-DACHEX)] derived from oxaliplatin. During the testing of the above mentioned substances we focused on the analysis of the antiproliferative action of these complexes and their mechanism of action. The results of the work show that the R,R-enantiomer has high therapeutic effects when it is applied to the PSN1 pancreatic adenocarcinoma cells. The experiments also proved that the mechanism of action of this complex in pancreatic cancer cells involves influencing the lipogenesis pathway, namely inhibition of de novo lipid synthesis. The antitumor action aimed at influencing the metabolism represents a new mechanism of action that has not yet been considered for clinically used antitumor platinum drugs. The obtained information suggests that R,R-enantiomer could represent a future promising cytostatic that would cause fewer adverse effects on the patient.
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The role of phosphoinositides in macropinocytosis
Hájek, Tomáš ; Doubravská, Lenka (advisor) ; Španielová, Hana (referee)
Macropinocytosis is non-selective actin-dependent type of endocytosis. It is important for the normal physiology of some cell types. However, it is used by intracellular parasites which internalise themselves into host cells in this way and also play a role in the nutritional supply in some type of cancer cells. During macropinocytosis a self-organized subdomain of plasma membrane is separated by a diffusion barrier named macropinocytic cup. RAC1-driven actin polymerization is required for membrane protrusion at the cup periphery, where a narrow ring of the actin nucleating factors is present. In contrast, actin dissociation occurs at the base of the cup due to RAS-controlled formation of phosphatidylinositol trisphosphates (PIP3). During cup closure sequential breakdown of PIP3 to phosphatidylinositol and acquisition of the endosomal identity of the newly formed vesicle is necessary. As a result of tubulation in the early stages of macropinocytosome maturation the vesicle decreases in diameter and stabilizes. At late stages the macropinocytic vesicle may fuse with the lysosome, allowing internalized material to enter this degradative organelle. Throughout the process specific types of phosphatidylinositols are part of the membrane providing signal transduction and membrane identity. These phospholipids...
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Analysis of Microscopic Images of Cancer Cells
Vičar, Tomáš ; Matula,, Petr (referee) ; Sladoje, Natasa (referee) ; Kolář, Radim (advisor)
Tato disertační práce je zaměřena na analýzu různých forem mikroskopických obrazových dat nádorových buněk (statické 2D snímky, statické 3D obrazy, 2D časosběrné zobrazování živých buněk). Hlavní pozornost je věnována datům získaným koherencí řízeným holografickým mikroskopem, který je relativně novou modalitou schopnou kotrastních záznamů živých buněk bez barvení (label-free) a poskytuje kvantitativní informaci (kvantitativní fázové zobrazení - QPI). V práci je popsán základní postup analýzy těchto snímků a jsou vytvářeny nové metody a zdokonalovány metody pro jednotlivé kroky této analýzy. Největší část práce je věnována segmentaci buněk, kde jsou shrnuty klasické metody i metody založené na hlubokém učení. Jsou také vyvinuty nové metody vhodné právě pro QPI data. Část práce je také věnována segmentaci 3D fluorescenční jader a detekci DNA zlomů pomocí hlubokého učení. Práce se zabývá i dalším zpracování v podobě sledování buněk, extrakce příznaků a následné analýze, kde je detekována buněčná smrt a jsou vytvořeny vhodné interpretovatelné příznaky pro klasifikaci buněčné smrti na apoptickou a lytickou. Celkově tato práce přispívá k rozvoji jednotlivých kroků analýzy obrazu nádorových buněk a odráží současný pokrok v oblasti analýzy obrazu, zejména přístupy hlubokého učení, což je také demonstrováno na několika výzkumných aplikacích.
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Analysis of changes in the phenotype of tumour cells induced by migrastatics in quantitative phase imaging
Kolínková, Veronika ; Netíková,, Irena Štenglová (referee) ; Veselý, Pavel (advisor)
This thesis focuses on the observation of living cells using the non-invasive methods of quantitative phase imaging (hiQPI). The imaging is enabled by a coherence-controlled holographic microscope (CCHM) developed in the Laboratory of Experimental Biophotonics at the VUT. Using this imaging technology, morphological changes of A549 and MCF7 after application of potential migrastatic drugs tumor cells are evaluated in the experimental part of the thesis. Migrastatics are defined as already approved drugs that could prevent the migration of cancer cells from the primary tumor and thus prevent metastasis. The RAC-GM (Rapid Assessment of Cell Growth and Migration in Vitro) method was chosen to assess their effect on tumor cells.
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Effect of cancer-associated fibroblasts on the survival, proliferation and invasiveness of cancer cells.
Nováková, Gita ; Anděra, Ladislav (advisor) ; Brábek, Jan (referee)
Tumour microenvironment, in addition to cancer cells themselves, represents important structural and functional part of the tumour. Similarly to the normal organs tumour microenvironment comprises several cell types (fibroblasts, immune cells, endothelial cells etc.) and non-cellular components, particularly extracellular matrix. All of them form favourable conditions for the growth, proliferation, protection from the immune system- mediated destruction and nutrition of cancer cells. Cancer associated fibroblasts (CAFs) represent the most abundant cell type of tumour microenvironment. Their origin can be traced to local normal fibroblasts, endothelial cells or epithelial cells and the transition into the CAFs phenotype is influenced with several factors secreted by cancer cells (particularly TGF-β). In contrast to fibroblasts activated during wound healing newly formed cancer associated fibroblasts expressing α-SMA are not subsequently eliminated from the respektive tissue. They persist and produce a number of pro-tumorigenic factors - SDF-1, HGF, IGF-1, IL-6, VEGF, PDGF-C, TGF-β, MMPs etc. CAFs and their secreted factors target several signalling pathways enhancing basic characteristics of the tumour, so called Hallmarks of Cancer. Cancer associated fibroblasts promote proliferation and invasiveness of...
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The role of caspase 2 in apoptosis induction in tumor cells.
Schmiedlová, Martina ; Kovář, Jan (advisor) ; Horníková, Lenka (referee)
Within the cell, caspase-2 probably fulfills several functions. Caspase-2 can be involved in apoptosis induction, DNA repair as well as cell cycle regulation. Caspase-2 has the character of initiator and also executioner caspase. A stimulus for caspase 2 activation can be oxidative stress or DNA damage. Caspase-2 is activated by cleavadge during an interaction with protein complexes. One of protein complexes,i.e. PIDDosome, is made of protein PIDD, RAIDD and pro-caspase-2. Withine the PIDDosome, caspase-2 is activated. Activated caspase-2 occures in a short S form and in long L form. L form of caspase-2 has proapoptotic effects and S form of caspase-2 has antiapoptotic effects. Caspase-2S has been only detected on mRNA level but not on protein level. The main role of caspase-2L is apoptosis induction in normal and tumor cells. Caspase-2 in tumour cells is activated by extrinstic as well as intristic apoptotic pathway. Apoptosis induction by caspase-2 is for example studied in connection with breast cancer treatment with taxanes. Caspase-2 ability of apoptosis induction in cancer cells independently of p53 protein is employed in cancer treatment including overcoming the resistance to apoptosis induction which is based on loosing p53 activity. Caspase-2 is involved in apoptosis induction by different...
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