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Analysis of motility in leukemia cells using incoherent holographic quantitative phase imaging
Smrčková, Zuzana ; Škarková, Aneta (referee) ; Zicha, Daniel (advisor)
This diploma thesis deals with the issue of motility analysis in leukemia cells. An accurate description of the cell movement and the detection of differences in motility under experimental conditions can be obtained by quantitative analysis of cell motility using time-lapse recording. The first part of this work describes various types of tumor cell migraton. The second part focuses on methods of analysis of cell motility in tissue culture using time-lapse recording, which include image acquisition and processing. Part of this chapter describes a coherence-controlled holographic microscope, which was used in the practical part and for which an insert was designed to ensure the exact and stable position of the individual chambers. The last part is focused on the research of leukemic cell motility, which is concluded by a discussion of the obtained results. The appendix contains a published study included acknowledgement to the author of this diploma thesis for participation in the project.
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Watersoluble polymer nanomaterials tailored for anti-tumor therapy
Vinklerová, Laura ; Etrych, Tomáš (advisor) ; Škarková, Aneta (referee)
Anti-tumor treatment involves several therapeutic approaches. One of them is chemotherapy which uses low-molecular-weight cytostatic drugs, whose disadvantage is the systemic manifestation of cytostatic effects even in healthy tissue. As a result, at the end of the last century, a new concept of high molecular weight polymer nanomaterials was introduced that minimized the side effects of treatment. The binding of the drug to the polymeric nanomaterial makes it possible to improve the biodistribution of the drug in the body and thereby reduce its toxicity, while often leading to a significant increase in anti-tumor activity. The structure of the high-molecular-weight polymer nanomaterials and their drug conjugates utilizes the differences between healthy and tumor tissue. One of the important difference is the production of matrix metalloproteinase enzymes in the tumor microenvironment, which is mainly used to release polymer-bound drug in tumor tissue. Keywords: polymer nanomaterial, matrix metalloproteinase, release of the drug, penetration, EPR efect
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The analysis of cancer invasion plasticity in a 3D environment
Škarková, Aneta ; Brábek, Jan (advisor) ; Anděra, Ladislav (referee) ; Bryja, Vítězslav (referee)
iii Abstract Cells have evolved multiple mechanisms of cellular motility ranging from the migration of large cell cohorts to specialized migration of individual cells. The wide range of invasion modes has been exploited by cancer cells to their advantage, which has rendered the metastatic process so difficult to defeat. To allow for a better understanding of cancer invasion plasticity, we have employed studies on cancer cells that adopt the proteolytically active, adhesion-dependent, elongated mesenchymal invasion mode, the protease-independent, low adhesion, rounded amoeboid invasion mode, or combination of both. To study invasion plasticity directly, we have established two model systems of the mesenchymal- amoeboid transition (MAT) that allow for regulated induction of MAT in 3D in vitro environments. Using these systems, MAT was induced in HT1080 fibrosarcoma cells and the acquisition of a motile, invasive amoeboid phenotype was confirmed. We then observed the mesenchymal and amoeboid invasion strategies within 3D collagen in more detail using a digital holographic microscope. Further, HT1080 cells before and after MAT were subject to high throughput proteomic and transcriptomic studies. Comparison of gene expression and protein levels of mesenchymal and amoeboid cells disclosed an inflammatory-like...
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Pathologic STAT3 signalling pathway activation in cancer and viral diseases.
Podestátová, Barbora ; Reiniš, Milan (advisor) ; Škarková, Aneta (referee)
STAT3, one of the seven members of STAT protein family, is able to transduce signal into the nucleus, where it binds to specific DNA sequences and acts as a transcription factor. Under physiological conditions, STAT3 regulates genes associated with number of functions such as cell proliferation, differentiation, apoptosis or immune response. In the case of pathological conditions, STAT3 can be dysregulated or constitutively activated, which may result in cancerogenesis. During this process, STAT3 is frequently activated directly in tumor cells where it acts tumorigenically. STAT3 is also associated with inflammatory reactions mediated by immune cells, which along with tumor and stromal cells are involved in the formation of the tumor microenvironment. The role of STAT3 is also important in the fight against viral infections, and when STAT3 activated aberrantly, it can lead to chronic diseases, including cancer. Due to these serious roles during pathogenesis, STAT3 is the subject of research of various inhibitors that either directly inhibit the STAT3 molecule function or indirectly any of the components of its signaling pathway.
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Analysis of motility in leukemia cells using incoherent holographic quantitative phase imaging
Smrčková, Zuzana ; Škarková, Aneta (referee) ; Zicha, Daniel (advisor)
This diploma thesis deals with the issue of motility analysis in leukemia cells. An accurate description of the cell movement and the detection of differences in motility under experimental conditions can be obtained by quantitative analysis of cell motility using time-lapse recording. The first part of this work describes various types of tumor cell migraton. The second part focuses on methods of analysis of cell motility in tissue culture using time-lapse recording, which include image acquisition and processing. Part of this chapter describes a coherence-controlled holographic microscope, which was used in the practical part and for which an insert was designed to ensure the exact and stable position of the individual chambers. The last part is focused on the research of leukemic cell motility, which is concluded by a discussion of the obtained results. The appendix contains a published study included acknowledgement to the author of this diploma thesis for participation in the project.
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Watersoluble polymer nanomaterials tailored for anti-tumor therapy
Vinklerová, Laura ; Etrych, Tomáš (advisor) ; Škarková, Aneta (referee)
Anti-tumor treatment involves several therapeutic approaches. One of them is chemotherapy which uses low-molecular-weight cytostatic drugs, whose disadvantage is the systemic manifestation of cytostatic effects even in healthy tissue. As a result, at the end of the last century, a new concept of high molecular weight polymer nanomaterials was introduced that minimized the side effects of treatment. The binding of the drug to the polymeric nanomaterial makes it possible to improve the biodistribution of the drug in the body and thereby reduce its toxicity, while often leading to a significant increase in anti-tumor activity. The structure of the high-molecular-weight polymer nanomaterials and their drug conjugates utilizes the differences between healthy and tumor tissue. One of the important difference is the production of matrix metalloproteinase enzymes in the tumor microenvironment, which is mainly used to release polymer-bound drug in tumor tissue. Keywords: polymer nanomaterial, matrix metalloproteinase, release of the drug, penetration, EPR efect
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