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Uloha kataninu, ATPázy štěpící mikrotubuly, při modulaci buněčné motility a proliferace glioblastomových buněk.
Uhlířová, Jana ; Dráber, Pavel (advisor) ; Libusová, Lenka (referee)
Glioblastomas are the most common and the deadliest types of brain tumours. Due to their highly invasive behaviour, they are incurable by convencial therapeutical strategies. It was shown that some components of microtubules, namely class III β-tubulin, γ-tubulin and microtubule severing protein spastin are overexpressed in glioblastoma cell lines as well as glioblastomas. This diploma thesis is focused on the expression, subcellular distribution and function of katanin, another microtubule-severing enzyme, in gliobastoma cell lines. Katanin is formed by catalytic (p60) and regulatory (p80) subunits. Expression and cellular localization of both katanin subunits was studied in panel of human glioblastoma cell lines isolated form adults (T98G, U87MG, U118MG and U138 MG) and child (KNS42). Data presented in this thesis demonstrated that katanin subunits were overexpresed both on transcript and protein levels in T98G, U87MG and KNS42 cell lines, but not in U138MG and U118MG cell lines when compared to normal non- transformed human astrocytes. Immunofluorescence microscopy revealed that both katanin subunits were diffusively distributed in cytoplasm and concentrated on spindle poles of mitotic cells and on leading edges of migrating cells. Examination of cell motility revealed that velocities in...
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Uloha kataninu, ATPázy štěpící mikrotubuly, při modulaci buněčné motility a proliferace glioblastomových buněk.
Uhlířová, Jana ; Dráber, Pavel (advisor) ; Libusová, Lenka (referee)
Glioblastomas are the most common and the deadliest types of brain tumours. Due to their highly invasive behaviour, they are incurable by convencial therapeutical strategies. It was shown that some components of microtubules, namely class III β-tubulin, γ-tubulin and microtubule severing protein spastin are overexpressed in glioblastoma cell lines as well as glioblastomas. This diploma thesis is focused on the expression, subcellular distribution and function of katanin, another microtubule-severing enzyme, in gliobastoma cell lines. Katanin is formed by catalytic (p60) and regulatory (p80) subunits. Expression and cellular localization of both katanin subunits was studied in panel of human glioblastoma cell lines isolated form adults (T98G, U87MG, U118MG and U138 MG) and child (KNS42). Data presented in this thesis demonstrated that katanin subunits were overexpresed both on transcript and protein levels in T98G, U87MG and KNS42 cell lines, but not in U138MG and U118MG cell lines when compared to normal non- transformed human astrocytes. Immunofluorescence microscopy revealed that both katanin subunits were diffusively distributed in cytoplasm and concentrated on spindle poles of mitotic cells and on leading edges of migrating cells. Examination of cell motility revealed that velocities in...
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The role of microtubules splitting proteins in regulation of cytoskeleton.
Uhlířová, Jana ; Dráber, Pavel (advisor) ; Libusová, Lenka (referee)
Microtubules are highly dynamic cytoskeletal structures, which secure cell shape, intracellular transport and organization of cellular organelles. They also participate in cell migration and in formation of mitotic spindle. Microtubules create a highly organized network which changes its organisation in response to the immediate need of the cell. They are formed of αβ-tubulin dimers and a large number of associated proteins. Microtubules are a polar structures with (+) and (-) ends, that differ in the rate of assembly. For fast regulation of microtubule structure are important microtubule-severing proteins (spastin, katanin and fidgetin), that belong to the AAA ATPases. These proteins associate with microtubules and consequently sever microtubules into two parts creating new (+) and (-) ends. In this way, they regulate many cellular processes, such as maintaining of cell shape, cell motility, proliferation and differentiation. Beside microtubule-severing activity, spastin and fidgetin are involved in processes in cell nucleus. Spastin is also associated with membranes and participates in modulation of endoplasmic reticulum and generation of endosomal vesicles. Mutations of microtubule-severing proteins cause embryonal development disorders, nervous system defects or infertility. The latest research...
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