The effects of ascorbic acid on viability and metabolism of osteosarcoma stem cells Utjecaj askorbinske kiseline na vijabilnost i metabolizam matičnih stanica osteosarkoma

master's thesis

• Title (croatian): TUtjecaj askorbinske kiseline na vijabilnost i metabolizam matičnih stanica osteosarkoma
• Author: Lucija Librenjak
• Mentor: Inga Urlić (mentor)
• Committee member: Inga Urlić (predsjednik povjerenstva)
• Committee member: Petra Peharec Štefanić (član povjerenstva)
• Committee member: Domagoj Đikić (član povjerenstva)
• Committee member: Romana Gračan (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2021-07-16, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Osteosarcoma is the most common primary bone malignancy and is the leading cause of tumor-induced death in children and young adults. Due to the absence of specific symptoms, this tumor is often diagnosed after it has already metastasized. Osteosarcoma is highly invasive and often shows chemoresistance. Many studies suggest that cancer stem cells play a critical role in these deadly mechanisms of cancer. Although infrequent, but with the ability to self-renew and differentiate into different cell types, this population of tumor cells supports tumor growth and heterogeneity. To successfully eradicate the whole tumor and reduce the risk of disease recurrence, anticancer therapy should also target cancer stem cells. Apart from its antioxidant properties and depending on applied concentration, ascorbic acid can also behave as a pro-oxidant and cause metabolic stress of the cell. The general aim of this thesis was to investigate the effects of ascorbic acid on viability and metabolism of osteosarcoma stem cells. We isolated cancer stem cells from the patient’s biopsies using the sphere culture system and identified them as stem cells using the Hoechst 33342 exclusion assay. The cytotoxicity of high-dose ascorbic acid was confirmed by the MTT test, with osteosarcoma stem cells being the most sensitive. The Seahorse XF method showed that ascorbic acid reduces the glycolytic and aerobic metabolic potential of the cell.

Abstract (croatian)

Osteosarkom predstavlja najčešću primarnu malignost kosti i vodeći je uzrok smrti uzrokovanih tumorima kod djece i mladih odraslih. Zbog izostanka specifičnih simptoma ovaj se tumor često dijagnosticira nakon što je već metastazirao. Visoka stopa invazivnosti i kemorezistencije karakteristični su za osteosarkome. Mnoga istraživanja ukazuju na tumorske matične stanice kao potencijalne krivce i inicijatore tih smrtonosnih mehanizama tumora. Iako malobrojna, ova populacija stanica unutar tumorske mase svojom sposobnošću samoobnavljanja i diferencijacije u različite stanične tipove, podupire rast i heterogenost tumora. Uspješna terapija za potpunu eliminaciju tumora i smanjenje mogućnosti relapsa, trebala bi ciljati i tumorske matične stanice. Ovisno o koncentraciji, osim antioksidativnog, poznato je i prooksidativno djelovanje askorbinske kiseline koje može rezultirati metaboličkim stresom stanice. Glavni cilj ovog diplomskog rada bio je istražiti utjecaj askorbinske kiseline na vijabilnost i metabolizam matičnih stanica osteosarkoma. Iz biopsija zahvaćene kosti izolirali smo tumorske matične stanice koristeći metodu uzgoja stanica u sferama te smo dokazali njihovu matičnost pomoću metode izbacivanja Hoechst 33342 boje. Citotoksičnost visokih doza askorbinske kiseline potvrđena je MTT testom pri čemu su se najosjetljivijima pokazale upravo matične stanice osteosarkoma. Metoda Seahorse XF pokazala je da askorbinska kiselina smanjuje glikolitički i aerobni metabolički potencijal stanice.

• Keywords: oncology; cytotoxicity; Seahorse XF; energy phenotype; metabolic potential
• Keywords (croatian): onkologija; citotoksičnost; Seahorse XF; energetski fenotip; metabolički potencijal
• Language: croatian
• URN:NBN: urn:nbn:hr:217:131362
• 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: 2023-07-16
• Created on: 2021-07-22 13:12:21