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National Repository of Grey Literature 2 records found  Search took 0.01 seconds. 
The role of the WASH complex in endolysosomal homeostasis
 Beránková, Pavla ; Libusová, Lenka (advisor) ; Rösel, Daniel (referee)
The WASH (WASP and SCAR homologue) complex is an actin nucleation promoting factor essential for endosomal cargo sorting. Upon WASH complex depletion, endosomal cargoes are mislocalized and the endolysosomal system collapses. Here, we employed high-speed vesicle tracking and real-time rescue experiments to test the effect of WASH complex depletion on endolysosomal homeostasis. We found that large lysosome-like vacuoles emerge in knockout cell lines of individual WASH complex subunits, although the overall dynamics of the lysosomal network does not substantially change. Follow-up experiments revealed that the WASH complex does not act directly on the vacuoles during their rescue. Overall, the data indicate that the emergence of vacuoles in WASH complex knockouts is a secondary process that depends on the WASH complex indirectly.
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Potentation of the effect of cytostatic by v-ATPase inhibitors
 Vaňková, Kristýna ; Eckschlager, Tomáš (advisor) ; Hudeček, Jiří (referee)
Tumour diseases belong among some of the most severe problems of advanced civilization and of current medicine. Neuroblastoma is a solid embryonal children's tumour, which develops from the cells of the neural crest. The increased expression of some oncogenes and their mutation leading to dysfunction plays a major role in the pathogenesis of this disease. One of the possible treatments of the tumours is the use of cytostatics, substances with no selectivity, which kills both damaged and healthy cells by their toxic effect. An example of such a cytotoxic substance is cisplatin (CDDP) which binds with purine nucleotides and leads the cell to apoptosis. A current complication in the treatment is development of chemoresistance. In connection with this issue vacuolar-type H+ -ATPase (V-ATPase) responsible for the acidification of vacuoles is studied. This bachelor thesis concentrates on studying the expression of selected genes of the V- ATPase in the neuroblastoma stem cell line sensitive (UKF-NB-4) and resistant to cisplatin (UKF-NB-4CDDP) and after their exposure to cisplatin. That was accomplished on the genetic level by the isolation of cellular RNA, reverse transcription creating complementary DNA (cDNA) and a method of a real time quantitative polymerase chain reaction (RT-PCR). The aim of the...
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