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master's thesis
Kloniranje urokinaznog plazminogenskog aktivatora, njegovog inhibitora i receptora te njihova ekspresija u ljudskim stanicama u kulturi
Zagreb: University of Zagreb, Faculty of Science, 2022. urn:nbn:hr:217:049696
Balent, Tihana
University of Zagreb
Faculty of Science
Department of Biology

Institutional repository: Repository of the Faculty of Science

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Balent, T. (2022). Kloniranje urokinaznog plazminogenskog aktivatora, njegovog inhibitora i receptora te njihova ekspresija u ljudskim stanicama u kulturi (Master's thesis). Zagreb: University of Zagreb, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:217:049696

Balent, Tihana. "Kloniranje urokinaznog plazminogenskog aktivatora, njegovog inhibitora i receptora te njihova ekspresija u ljudskim stanicama u kulturi." Master's thesis, University of Zagreb, Faculty of Science, 2022. https://urn.nsk.hr/urn:nbn:hr:217:049696

Balent, Tihana. "Kloniranje urokinaznog plazminogenskog aktivatora, njegovog inhibitora i receptora te njihova ekspresija u ljudskim stanicama u kulturi." Master's thesis, University of Zagreb, Faculty of Science, 2022. https://urn.nsk.hr/urn:nbn:hr:217:049696

Balent, T. (2022). 'Kloniranje urokinaznog plazminogenskog aktivatora, njegovog inhibitora i receptora te njihova ekspresija u ljudskim stanicama u kulturi', Master's thesis, University of Zagreb, Faculty of Science, accessed 04 November 2024, https://urn.nsk.hr/urn:nbn:hr:217:049696

Balent T. Kloniranje urokinaznog plazminogenskog aktivatora, njegovog inhibitora i receptora te njihova ekspresija u ljudskim stanicama u kulturi [Master's thesis]. Zagreb: University of Zagreb, Faculty of Science; 2022 [cited 2024 November 04] Available at: https://urn.nsk.hr/urn:nbn:hr:217:049696

T. Balent, "Kloniranje urokinaznog plazminogenskog aktivatora, njegovog inhibitora i receptora te njihova ekspresija u ljudskim stanicama u kulturi", Master's thesis, University of Zagreb, Faculty of Science, Zagreb, 2022. Available at: https://urn.nsk.hr/urn:nbn:hr:217:049696

Metadata
TitleKloniranje urokinaznog plazminogenskog aktivatora, njegovog inhibitora i receptora te njihova ekspresija u ljudskim stanicama u kulturi
Title (english)Cloning of urokinase plasminogen activator, its inhibitor and receptor and their expression in human cells in culture
AuthorTihana Balent
MentorMaja Matulić (mentor)
Committee memberMaja Matulić (predsjednik povjerenstva)
Committee memberŽeljka Soldin (član povjerenstva)
Committee memberInes Radanović (član povjerenstva)
Committee memberDraginja Mrvoš-Sermek (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Science
(Department of Biology)
Zagreb
Defense date and country2022-02-25, Croatia
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Biology
Abstract
Urokinazni plazminogenski aktivator (uPA) ili urokinaza zaslužan je za aktivaciju plazminogena i njegovu pretvorbu u plazmin, proteolitički enzim koji razgrađuje ekstracelularni matriks. Urokinaza je dio sustava koji obuhvaća i receptor uPA (uPAR) koji je veže na staničnu membranu te njene inhibitore, PAI-1 i PAI-2. Cilj ovog rada bio je klonirati sekvence koje kodiraju za urokinazu, njen receptor i inhibitor PAI-1 te ih eksprimirati u različitim staničnim linijama. Analiza ekstracelularne enzimske aktivnosti i ekspresije pojedinih elemenata sustava u različitim tipovima stanica ukazala bi na mehanizme regulacije urokinazne aktivnosti. Konstruirani su plazmidi sa sekvencama gena za uPA, PAI-1 i uPAR te transficirane stanične linije A1235, HeLa i HEK293. Stanice HEK293 i A1235 transficirane plazmidom sa sekvencom za uPA imale su bazalnu aktivnost urokinaze. Sojevi stanica A1235 i HEK293 transficirani plazmidom sa sekvencom za PAI-1, pokazali su smanjenu aktivnost urokinaze. Stanice HeLa nisu pokazale urokinaznu aktivnost. Unos plazmida sa sekvencom uPAR nije utjecao na urokinaznu aktivnost stanica. Analiza ekspresije proteina pokazala je da plazmidi sa sekvencom urokinaze nisu promijenili razinu ekspresije ovog proteina u staničnim lizatima. Kod stanica A1235 i HEK293 detektirano je povećanje ekspresije uPAR i PAI-1 nakon transfekcije plazmidima s njihovim sekvencama. Uzgojene stanične linije s različitom urokinaznom aktivnosti mogu poslužiti kao model za istraživanje utjecaja urokinaznog sustava na staničnu proliferaciju, migraciju i preživljenje.
Abstract (english)
Urokinase plasminogen activator (uPA) or urokinase is responsible for the activation of plasminogen and its conversion into plasmin, a proteolytic enzyme that degrades the extracellular matrix. Urokinase is a part of a system that includes the uPA receptor (uPAR) that binds urokinase to the cell membrane and its inhibitors, PAI-1 and PAI-2. The aim of this study was to clone sequences encoding urokinase, its receptor and PAI-1, and to express them in different cell lines. Analysis of extracellular enzyme activity and expression of individual urokinase system elements in different cell types, could indicate the regulatory mechanisms of urokinase activity. Plasmids with gene sequences for uPA, PAI-1 and uPAR were constructed and A1235, HeLa and HEK293 cell lines were transfected with plasmids. HEK293 and A1235 cells transfected with the uPA sequence plasmid had basal urokinase activity. A1235 and HEK293 cell strains transfected with plasmid containing PAI-1 sequence showed reduced urokinase activity. HeLa cells did not have urokinase activity. Expression of plasmids with the uPAR sequence did not affect urokinase activity in the transfected cell lines. Analysis of protein expression showed that plasmids with the urokinase sequence did not change the expression levels of this protein in the cell lysates. In A1235 and HEK293 cells, an increase in uPAR and PAI-1 expression was detected after transfection with plasmids with corresponding sequences. Cell strains with different urokinase activity can serve as a model for investigating the impact of the urokinase system on cell proliferation, migration and survival.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:217:049696
Study programmeTitle: Integrated undergraduate and graduate university study of Biology and Chemistry Education; specializations in: Integrated undergraduate and graduate university study of Biology and Chemistry Education
Course: Integrated undergraduate and graduate university study of Biology and Chemistry Education
Study programme type: university
Study level: integrated undergraduate and graduate
Academic / professional title: magistar/magistra edukacije biologije i kemije (magistar/magistra edukacije biologije i kemije)
Type of resourceText
File originBorn digital
Access conditionsOpen access
Embargo expiration date: 2024-02-25
Terms of useLicence In copyright icon
Created on2022-03-23 13:23:49