Analiza antiproliferativnog učinka Schwannovih stanica na stanice neuroblastoma

master's thesis

• Title (english): Analysis of the antiproliferative and differentiation-inducing effects of the Schwann cells on neuroblastoma cells
• Author: Helena Dodig
• Mentor: Peter Ambros (mentor)
• Mentor: Petra Korać (mentor)
• Committee member: Petra Korać (predsjednik povjerenstva)
• Committee member: Vesna Benković (član povjerenstva)
• Committee member: Damjan Franjević (član povjerenstva)
• Committee member: Inga Marijanović (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2017-05-19, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Schwannove su stanice neophodne za proces sazrijevanja benignih tipova neuroblastoma, ganglioneuroma, djelujući kroz neuritogenične, anti-proliferativne i pro-apoptostske signale. Cilj ovog diplomskog rada bio je identificirati proteine koje izlučuju Schwanove stanice, a koji sudjeluju u njihovoj interakciji sa stanicama neuroblastoma, te definirati koncentraciju i kombinaciju potrebnu za inhibiciju rasta stanica neuroblastoma. Usporedbom rezultata dobivenih pomoću analize proteina i podataka dobivenih sekvenciranjem transkriptoma s podacima iz literature, izabrano je osam faktora: IGFBP6, FGF7, CNTF, PTN, NGF, BDNF, GDNF i EGFL8. Za funkcionalnu potvrdu njihova djelovanja in vitro, odgovarajući su rekombinantni proteini dodani u medij korišten za kultiviranje dviju staničnih linija neuroblastoma porijeklom iz agresivnih tumora, STA-NB-6 i STA-NB-10, te je učinak izmjeren pomoću protočne citometrije. Ovo je istraživanje identificiralo PTN, IGFBP6 i EGFL8 kao proteine koje izlučuju Schwannove stanice, koji, kada su individualno dodani staničnom mediju tijekom 17 dana, smanjuju proliferaciju i potiču diferencijaciju STA-NB-6 stanica. NGF je potvrđen kao neuritogenični faktor STA-NB-6 stanica. Kombinacija svih navedenih faktora nije imala značajni učinak na STA-NB-6 stanice. Faktori nisu imali značajan učinak na STA-NB-10 stanice. Ovaj rad daje osnovu za daljnja in vitro i buduća istraživanja in vivo, za ispitivanje učinka navedenih faktora na rast neuroblastoma metodom staničnog eksplantata.

Abstract (english)

Schwann cells are essential for the maturation process of benign forms of neuroblastoma, ganglioneuroma, acting via neuritogenic, anti-proliferative and pro-apoptotic signals. The goal of this master thesis was to identify Schwann cell-secreted proteins involved in the cross-talk with neuroblastoma cells, and define the concentration and combination needed to induce a growth-impairing effect in neuroblastoma cells. Eight factors were chosen based on protein array results, RNA-sequencing data and literature research: IGFBP6, FGF7, CNTF, PTN, NGF, BDNF, GDNF and EGFL8. They were functionally validated in vitro by cultivating two neuroblastoma cell lines derived from aggressive tumors, STA-NB-6 and STA-NB-10, in presence of corresponding recombinant proteins. The effects were measured by flow cytometry. PTN, IGFBP6 and EGFL8 were identified as Schwann cell-secreted proteins that cause reduction of cell proliferation and induction of neuronal-like differentiation of STA-NB-6 cells, when individually added to culture media for 17 days. NGF was confirmed as a neuritogenic factor of STA-NB-6 cells. The combination of all factors had no significant effect on STA-NB-6 cells. The factors had no significant effect on STA-NB-10 cells. This work forms basis for further in vitro and in vivo experiments to address the effect of these factors on neuroblastoma growth in xenograft models.

• Keywords: Schwannove stanice; neuroblastom; proteinska rešetka; sekvenciranje transkriptoma; protočna citometrija; stanična proliferacija; stanična diferencijacija
• Keywords (english): Schwann cells; neuroblastoma; protein array; RNA sequencing; flow cytometry; cell proliferation; cell differentiation
• Language: croatian
• URN:NBN: urn:nbn:hr:217:328200
• 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; Embargo expiration date: 2019-05-19
• Created on: 2017-05-26 08:25:44