Vloga proteina MAGE C2 pri adheziji celic glioblastoma

Banovšek, Nika (author), BREZNIK, BARBARA (mentor)

Glioblastom (GBM) je najpogostejši in najagresivnejši primarni možganski tumor pri odraslih. Zaradi svoje heterogenosti in pospešene invazije celic v centralnem živčnem sistemu je odporen na obstoječe oblike zdravljenja, kot so kirurška odstranitev, kemoterapija in radioterapija. Zaradi slabe prognoze želimo bolje razumeti napredovanje GBM in najti biološke označevalce in tarče, s katerimi bi izboljšali obravnavo bolnikov. Družino genov z melanomom povezanih antigenov (angl. melanoma associated antigen, MAGE) delimo na tip 1 in 2. Zadnje raziskave kažejo, da MAGE tipa 1 v različnih kombinacijah zagotavljajo zaščito pred različnimi stresnimi dejavniki, potencialno tudi zaščito rakavih celic pred zdravljenjem s kemoterapijo in z obsevanjem. Namen magistrskega dela je bil ugotoviti vpliv obsevanja in MAGE C2 na viabilnost, proliferacijo in adhezijo celic GBM. Obenem smo želeli oceniti njun vpliv na izražanje različnih adhezijskih molekul (CAM), proteinov zunajceličnega ogrodja (ECM) in citoskeleta, ki igrajo ključno vlogo pri napredovanju GBM. Uporabili smo celični liniji GBM LN-229 s stabilnim izražanjem proteina MAGE C2 in kontrolni celični liniji brez izraženega MAGE C2. Vpliv obsevanja in izražanje MAGE C2 smo analizirali s štetjem viabilnih celic ter kvantifikacijo adhezije celic po štiridnevnem frakcioniranem obsevanju z 2,5 Gy. S pomočjo prenosa po westernu in imunodetekcijo smo opazovali, kako obsevanje in MAGE C2 vplivata na izražanje proteinov CAM, ECM in citoskeleta. Naše ugotovitve kažejo, da frakcionirano obsevanje zmanjša proliferacijo in povečajo adhezijo celic GBM. Prav tako je obsevanje v celicah vplivalo na zmanjšanje izražanja celokupnega N-kadherina in upad alfa1-verige glede na celokupen kolagen I in III. Izražanje MAGE C2 v celicah ni vplivalo na občutljivost celic GBM na obsevanje in na adhezijo celic, saj ni prišlo do spremenjene celične proliferacije po obsevanju ter celične adhezije v celicah z izraženim MAGE C2 v primerjavi s celicami brez MAGE C2. Celice z izraženim MAGE C2 so imele napram celicam brez izraženega MAGE C2 nižje nivoje izražanja celokupnega N-kadherina in nižje razmerje cepljene in celokupne oblike ter alfa1-verige in celokupne oblike kolagena I in III. Statistične analize prenosa po westernu so pokazale, da obsevanje in MAGE C2 vplivata na raven izražanja N-kadherina ter kolagena I in III. Rezultati nakazujejo, da MAGE C2 vpliva na izražanje proteinov ECM in CAM. MAGE C2 tako predstavlja potencialen dejavnik, ki lahko posredno vpliva na migracijo in invazijo celic GBM.

• Language: Slovenian
• Keywords: glioblastom, LN-229, MAGE C2, obsevanje, celična adhezija, proliferacija
• Work type: Master's thesis/paper
• Typology: 2.09 - Master's Thesis
• Organization: FKKT - Faculty of Chemistry and Chemical Technology
• Year: 2025
• PID: 20.500.12556/RUL-173298
• COBISS.SI-ID: 257286403
• Publication date in RUL: 15.09.2025

Secondary language

• Language: English
• Title: The role of MAGE C2 Protein in Glioblastoma Cell Adhesion

Abstract

Glioblastoma (GBM) is the most common and most aggressive primary brain tumor in adults. Due to its heterogeneity and accelerated invasion of cells into the central nervous system, it is resistant to existing treatments such as surgical removal, chemotherapy, and radiotherapy. Because of the poor prognosis, we aim to better understand glioblastoma progression and identify biological markers that could improve patient management. MAGE (melanoma associated antigen) gene family is divided into type 1 and 2. Recent studies show that type 1 MAGEs ensure protection from various stress factors, potentially including protection of tumor cells from chemotherapy and radiation treatment. The aim of this study was to investigate the effects of irradiation and MAGE C2 on cell viability, proliferation, adhesion, and the expression of different adhesion molecules (CAM), extracellular matrix (ECM) proteins, and cytoskeletal proteins, which play a crucial role in GBM progression. We used GBM LN-229 cell lines with stable expression of MAGE C2 protein and control cell lines without MAGE C2 expression. The effects of irradiation and MAGE C2 were analysed by counting viable cells and quantifying cell adhesion after four days of fractionated irradiation with 2,5 Gy. Using western blotting and immunodetection, we examined how irradiation and MAGE C2 affect the expression of CAMs, ECM proteins and cytoskeletal proteins. Our findings show that fractionated irradiation reduces proliferation and increases cell adhesion of GBM cells. Irradiation also reduced the expression of total N-cadherin and decreased the alpha1-chain relative to total collagen I and III. Expression of MAGE C2 in cells did not affect the sensitivity of GBM cells to irradiation or cell adhesion, as there were no changes in cell proliferation, or in adhesion in MAGE C2-expressing cells compared with cells without MAGE C2. Cells expressing MAGE C2 displayed lower levels of total N-cadherin expression and lower ratios of cleaved-to-total collagen I and III, as well as lower ratios of alpha1-chain-to-total collagen I and III. Statistical analysis of Western blot results demostrated that irradiation and MAGE C2 affect the expression levels of N-cadherin, as well as collagen I and III. Our results suggest that MAGE C2 influences the expression ECM proteins and CAMs. Thus, MAGE C2 represents a potential factor that may indirectly contribute to GBM cell migration and invasion.

• Keywords: glioblastoma, LN-229, MAGEC2, irradiation, adhesion, proliferation