Vpliv zunajceličnega matriksa na preživetje in invazijo sferoidov glioblastomskih celic tretiranih s temozolomidom : magistrsko delo

Kovačič, Tinkara

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Title:	Vpliv zunajceličnega matriksa na preživetje in invazijo sferoidov glioblastomskih celic tretiranih s temozolomidom : magistrsko delo
Authors:	ID Kovačič, Tinkara (Author) ID Potočnik, Uroš (Mentor) More about this mentor... ID Zottel, Alja (Comentor)
Files:	MAG_Kovacic_Tinkara_2025.pdf (3,40 MB) MD5: B33BDF02E3B963458B808976CA3016DE
Language:	Slovenian
Work type:	Master's thesis/paper
Typology:	2.09 - Master's Thesis
Organization:	FKKT - Faculty of Chemistry and Chemical Engineering
Abstract:	Glioblastom (GBM) je najpogostejši in najbolj agresiven primarni možganski tumor pri odraslih, ki zaradi izrazite heterogenosti, odpornosti na terapijo in visoke invazivnosti vodi v hitro napredovanje bolezni z nizko stopnjo preživetja bolnikov. Zdravljenje GBM obsega kirurško odstranitev tumorja, radioterapijo in kemoterapijo s temozolomidom (TMZ). Kljub zdravljenju večina bolnikov ne preživi več kot 2 leti po diagnozi, zato so raziskave na tem področju, ki bi vodile do boljšega preživetja bolnikov, nujno potrebne. Vendar pa za ustrezno testiranje učinkovin potrebujemo biološko relevantne modele, ki bi posnemali naravo tumorja in njegovo mikrookolje. V magistrskem delu smo ovrednotili vpliv zunajceličnega matriksa na preživetje in invazijo sferoidov glioblastomskih celic tretiranih s TMZ. Sferoidi namreč bolj realistično ponazarjajo značilnosti tumorja in vedenje glioblastomskih celic v primerjavi z 2D modeli. Uporabili smo sferoide treh različnih glioblastomskih celičnih linij, HROG02, HROG06 in HROG10, ki smo jih razvili z metodo uporabe plošč z ultranizko pritrditvijo celic. Sferoide smo nato vgradili v zunajcelični matriks, sestavljen iz kolagena ali Matrigela, dveh različnih gostot. Nato smo različne modele, 2D kulture, sferoide brez matriksa, sferoide v kolagenu (40 in 200 μg/mL) in sferoide v Matrigelu (5 % in 30 %) tretirali z različnimi koncentracijami TMZ. Po 72 urah smo z uporabo reagenta WST-1 določili preživetje celic in izračunali IC50. Hkrati pa smo določili invazijo posameznih sferoidov vklopljenih v matriks. Ugotovili smo, da so sferoidi HROG02 brez matriksa bolj odporni na TMZ kot 2D kulture, medtem ko pri ostalih dveh linijah ni bistvenih razlik. Rezultati preživetja pri najvišji testirani koncentraciji TMZ kažejo, da višja koncentracija kolagena prispeva k večji odpornosti sferoidov HROG02 in HROG06, medtem ko je bilo preživetje v obeh koncentracijah Matrigela primerljivo. Rezultati invazije kažejo, da je invazija sferoidov HROG02 in HROG10 v matriksu z višjo koncentracijo Matrigela večja, vpliv TMZ pa šibkejši v primerjavi z matriksom z nižjo koncentracijo Matrigela. TMZ ni vplival na invazijo HROG06. Na podlagi rezultatov lahko sklepamo, da je odziv sferoidov glioblastomskih celic na tretiranje s TMZ kompleksen in pogojen z arhitekturo modela, biološkimi lastnostmi celične linije ter tipom in koncentracijo zunajceličnega matriksa. Naše raziskave odpirajo možnosti za nadaljnji razvoj biološko relevantnih modelov GBM, ki bi lahko izboljšali razumevanje terapevtskih odzivov in prispevali k učinkovitejšemu zdravljenju.
Keywords:	glioblastom, 2D kulture, sferoidi, zunajcelični matriks, temozolomid
Place of publishing:	Maribor
Place of performance:	Maribor
Publisher:	[T. Kovačič]
Year of publishing:	2025
Number of pages:	1 spletni vir (1 datoteka PDF (X, 52 str.))
PID:	20.500.12556/DKUM-95889
UDC:	616.714.1-006(043.2)
COBISS.SI-ID:	261097475
Publication date in DKUM:	20.11.2025
Views:	194
Downloads:	98
Metadata:
Categories:	KTFMB - FKKT
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Licences

License:	CC BY-NC-ND 4.0, Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International

Link:	http://creativecommons.org/licenses/by-nc-nd/4.0/
Description:	The most restrictive Creative Commons license. This only allows people to download and share the work for no commercial gain and for no other purposes.
Licensing start date:	05.11.2025

Secondary language

Language:	English
Title:	The effect of extracellular matrix on survival and invasion of glioblastoma spheroids treated with temozolomide
Abstract:	Glioblastoma (GBM) is the most common and most aggressive primary brain tumor in adults. Due to its pronounced heterogeneity, therapy resistance, and high invasiveness, it leads to rapid disease progression and poor patient survival. Standard treatment includes surgical resection of the tumor, radiotherapy, and chemotherapy with temozolomide (TMZ). Despite intensive therapy, most patients do not survive more than two years after diagnosis, which highlights the urgent need for research aimed at improving patient outcomes. However, the development of biologically relevant models that mimic the nature of the tumor and its microenvironment is essential for reliable drug testing. In this Master’s thesis, we evaluated the effect of the extracellular matrix on the survival and invasion of glioblastoma cell spheroids treated with TMZ. Spheroids were used as they better represent the characteristics and behavior of glioblastoma cells compared to 2D models. We used three glioblastoma cell lines (HROG02, HROG06, and HROG10) grown using ultra-low attachment plates. The resulting spheroids were embedded in extracellular matrix composed of either collagen or Matrigel at two different concentrations. We treated various models – 2D cultures, spheroids without matrix, spheroids in collagen (40 and 200 μg/mL), and spheroids in Matrigel (5 % and 30 %) – with different concentrations of TMZ. After 72 hours, cell viability was assessed using the WST-1 reagent, and IC50 values were calculated. In parallel, we quantified the invasion of spheroids embedded in extracellular matrix. We found that HROG02 spheroids without extracellular matrix were more resistant to temozolomide (TMZ) compared to 2D cultures, whereas no major differences were observed for the other two cell lines. Survival results at the highest tested TMZ concentration indicate that a higher collagen concentration contributes to increased resistance of HROG02 and HROG06 spheroids, while survival in both Matrigel concentrations was comparable. Invasion analysis showed that the invasion of HROG02 and HROG10 spheroids was higher in matrix with higher Matrigel concentration, and the effect of TMZ was weaker compared to matrix with lower Matrigel concentration. TMZ had no detectable effect on the invasion of HROG06 spheroids. These findings suggest that the response of glioblastoma spheroids to TMZ is complex and influenced by model architecture, cell line-specific characteristics, and the type and concentration of extracellular matrix. Our research supports the further development of biologically relevant glioblastoma models, which could improve the understanding of therapeutic responses and contribute to more effective treatment strategies.
Keywords:	glioblastoma, 2D cell cultures, spheroids, extracellular matrix, temozolomide

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