RNA sequencig of glioblastoma stem cells after induction of ferroptosis
Patient derived glioblastoma stem like cells cultured in vitro were used to investigate the effect of hypoxia and ferroptosis induction on gene expression. Different baseline transcriptomic profiles of studied cells indicated their diverse niche origin within the tumor. Futher analysis revealed that intrinsic gene expression of cells were exceeding the effect of hypoxia or treatment. Response of the cells whose profiles were linked with occupancy of more periferal parts of tumor (leading edge) differed from those that were residing in the tumor core (peri-nectrotic zone). Expression of genes related to respone to hypoxia and ferroptosis indicated more adaptation and possible higher resistance of the cells from tumor core. Overall design: Glioblastoma stem like cells derived from patients (GSCs, n=8) treated with DMSO (0.1% final concentration), and ferroptosis inducer at 3 uM (RSL3 in DMSO). Subsequently cells were cultured for 24h in either Normoxia (21% oxygen and 5% CO2) or Hypoxia (1% oxygen, 5% CO2). Cells were centrifuged (500xg, 5 min), medium decanted and pellets snap frozen for RNA after isolation with phenol-chloroform method combined with isopropanol precipitation at -80C overnight. -------------- Funding: FBW-05/2022 IMDIK PAN, K. Krawczynski G-1252-2022/45/B/NZ5/04226 NCN, A. Bronisz