Utjecaj retinoične kiseline i inhibitora poli (ADP-ribozil) polimeraze-1 na stanične linije neuroblastoma

master's thesis

• Title (english): Effect of retinoic acid and poly(ADP-ribose) polymerase-1 inhibitor on neuroblastoma cell lines
• Author: Martina Grubar
• Mentor: Maja Matulić (mentor)
• Committee member: Maja Matulić (predsjednik povjerenstva)
• Committee member: Zlatko Liber (član povjerenstva)
• Committee member: Ana Galov (član povjerenstva)
• Committee member: Inga Marijanović (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2017-05-15, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Proces diferencijacije obrnuto je razmjeran staničnom proliferacijskom potencijalu. Kao posljedica te pojave, jedan od mehanizama tumorigeneze je upravo zaustavljanje diferencijacijskih procesa. Pojedine tumorske stanične linije u kulturi mogu se diferencirati, a jedna od tvari koja može potaknuti te procese je retinoična kiselina. U ovom radu diferencijacija se pratila na modelu stanica neuroblastoma koje su se obrađivale retinoičnom kiselinom kroz duži vremenski period. Dok je linija neuroblastoma SH-SY5Y imala sposobnost diferencijacije, druga, neuroglioblastomska linija H4, nije pokazivala morfološke promjene tijekom tretmana. Stanice su se tretirale retinoičnom kiselinom uz i bez prisustva inhibitora poli (ADP-ribozil) polimeraze (PARP-1), enzima koji sudjeluje u procesima remodeliranja kromatina. Cilj ovog istraživanja bio je utvrditi utjecaj inhibicije PARP-1 na diferencijaciju induciranu retinoičnom kiselinom kod stanica SH-SY5Y te analizirati utjecaj retinočne kiseline i PARP-1 na liniju H4. Mjerenjem proliferacije stanica, duljine neurita i analizom ekspresije gena uključenih u diferencijaciju pokazano je da PARP-1 ne utječe na proces diferencijacije stanica SH-SY5Y. Kod linije H4 utvrđeno je da retinoična kiselina povećava aktivnost urokinaze, enzima uključenog u epitelno-mezenhimsku tranziciju (EMT), dok inhibicija PARP-1 nema utjecaj na njenu aktivnost. Međutim, inhibicija PARP-1 povećala je ekspresiju gena elemenata urokinaznog sustava i signalnog puta TGF-β te krovnih transkripcijskih faktora epitelno-mezenhimske tranzicije (EMT). Stoga se može zaključiti da bi PARP-1 mogao sudjelovati u procesima reprogramiranja kod stanične linije neuroglioblastoma.

Abstract (english)

Differentiation is inversely proportional to cellular proliferation potential. Consequently, one of the mechanisms of tumorigenesis is differentiation arrest. Some tumor cell lines can be differentiated in the cell culture and one of the agents that can induce differentiation is retinoic acid. In this thesis, processes of differentiation were examined on neuroblastoma cell model, which was treated with retinoic acid for a longer period. While neuroblastoma SH-SY5Y cell line was able to differentiate, the other, neuroglioblastoma H4 line, did not show morphological changes during treatment. Cells were treated with retinoic acid in the presence or absence of poli (ADP-ribose) polymerase (PARP-1) inhibitor, inhbitor of an enzyme that facilitates chromatin remodeling. The aim of this study was to determine the effect of PARP-1 inhibition on retinoic acid induced differentiation of SH-SY5Y cells and analyze the effect of retinoic acid and PARP-1 on H4 line. By measuring cell proliferation, neurite length and by analysis of gene expression involved in differentiation, it was shown that PARP-1 had no effect on SH-SY5Y differentiation. In H4 line it was found that retinoic acid induced urokinase activity. Urokinase is a plasminogen activator, an enzyme which could be involved in epithelial-mesenchymal transition (EMT). PARP-1 inhibition had no effect on the enzyme activity. However, PARP-1 inhibition increased the expression of urokinase system and TGF-β signaling pathway components’ genes, as well as EMT master transcription factors’ genes. Therefore, it can be concluded that PARP-1 could take part in reprogramming neuroglioblastoma cell line.

• Keywords: Neuroblastom; retinoična kiselina; PARP-1; diferencijacija; epitelno-mezenhimska tranzicija
• Keywords (english): Neuroblastoma; retinoic acid; PARP-1; differentiation; epithelial-mezenchymal transition
• Language: croatian
• URN:NBN: urn:nbn:hr:217:528738
• Study programme: Title: Molecular Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2017-05-26 08:21:16