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Vpliv obsevanja na glioblastomske matične celice in mezenhimske matične celice v kokulturi
ID Ušeničnik, Urška (Author), ID Lah Turnšek, Tamara (Mentor) More about this mentor... This link opens in a new window, ID Breznik, Barbara (Co-mentor)

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Abstract
Glioblastom je najpogostejši in najagresivnejši možganski tumor. Za visoko odpornost na standardno zdravljenje so odgovorne glioblastomske matične celice (GMC) in njihova interakcija s tumorskim mikrookoljem, katerega del so tudi mezenhimske matične celice (MMC). Raziskave imajo nasprotujoče rezultate glede vpliva MMC na rast in invazijo tumorja, vendar imajo velik potencial kot vektorji pri dostavljanju terapevtikov v tumor. Vpliv ionizirajočega sevanja, ki se rutinsko uporablja za zdravljenje glioblastoma, na GMC in MMC smo določali v tridimenzionalnem (3D) modelu sferoidov monokultur in kokultur obeh vrst celic. Izvedli smo test klonogenosti, invazije in proliferacije z imunobarvanjem s protitelesi proti Ki-67 in spremljali celični cikel s Hoechst barvilom. Rezultati testa klonogenosti so pokazali, da so GMC odporne na sevanje za razliko od MMC. Rast sferoidov v obsevani kokulturi GMC z MMC se je zmanjšala v primerjavi z obsevano monokulturo GMC. Invazija MMC v monokulturi se je povišala po obsevanju, v kokulturi z GMC pa sevanje ni imelo vpliva na invazijo MMC. Ionizirajoče sevanje in prisotnost MMC v kulturi nista imela vpliva na invazijo GMC. V monokulturi so se GMC hitreje delile kot MMC, v kokulturi pa so MMC svojo proliferacijo pospešile. Obsevanje je znižalo proliferacijo MMC in GMC v kokulturi. Prisotnost GMC in ionizirajoče sevanje ni imelo vpliva na celični cikel MMC. Celični cikel obsevanih GMC se je v monokulturi ustavil v G2 fazi, v kokulturi z MMC pa v G1 fazi. Naši rezultati so pripomogli k boljšem razumevanju odziva GMC na ionizirajoče sevanje in vpliv MMC na odpornost GMC na ionizirajoče sevanje. Rezultati so dobra iztočnica za nadaljne raziskave mehanizmov odpornosti glioblastoma na standardno zdravljenje.

Language:Slovenian
Keywords:glioblastom, glioblastomske matične celice, mezenhimske matične celice, kokultura, radioterapija, ionizirajoče sevanje, invazija, proliferacija, celični cikel
Work type:Master's thesis/paper
Typology:2.09 - Master's Thesis
Organization:BF - Biotechnical Faculty
Place of publishing:Ljubljana
Publisher:[U. Ušeničnik]
Year:2022
PID:20.500.12556/RUL-135556 This link opens in a new window
UDC:606:616-006:602.9:613.648.4(043.2)
COBISS.SI-ID:101355011 This link opens in a new window
Publication date in RUL:19.03.2022
Views:709
Downloads:59
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Secondary language

Language:English
Title:Effect of ionization on glioblastoma stem cells and mesenchymal stem cells in coculture
Abstract:
Glioblastoma is the most aggressive brain tumour. Its resistance to standard therapy is due to glioblastoma stem cells (GSC) and their interaction with tumour microenvironment, part of which are also mesenchymal stem cells (MSC). Research on MSC effects on tumour growth and invasion shows seemingly contradictory results. Still, MSC have great potential as vectors for drug delivery to the tumour. Here, we studied how ionizing radiation affects GSC and MSC in monocultures and cocultures, both in tridimensional (3D) spheroid model. We used clonogenic assay, invasion assay, proliferation assay with antibodies for Ki-67 protein and we also analysed cell cycle with Hoechst dye. GSC were more clonogenic then MSC, when irradiated. When in coculture with MSC, GSC spheroid growth slowed down compared to GSC monocultures. When irradiated, MSC were more invasive in monoculture than in coculture. Irradiation and MSC didn't influence invasion of GSC. GSC were more proliferative then MSC in monoculture. Proliferation of MSC increased in coculture but decreased when they were exposed to radiation. Proliferation of GSC decreased in irradiated coculture with MSC compared to radiated monoculture of GSC. GSC and radiation didn't affect cell cycle of MSC. When we radiated GSC in monoculture cell cycle stopped in G2 phase, but not when they were irradiated in the presence of MSC. Our results contribute to understanding of effect of irradiation on GSC and effect of MMC on GSC radioresistance. Results of this study are useful for future research of mechanisms that are responsible for glioblastoma resistance to standard therapy.

Keywords:glioblastoma, glioblastoma stem cells, mesenchymal stem cells, coculture, radiotherapy, ionizing radiation, invasion, proliferation, cell cycle

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