Activation of ferroptosis through GCL inhibition in chordoma

To evaluate the response of chordoma cell lines to ferroptosis activation through GCL inhibition, cell lines UM-Chor1, U-CHCF365, MUG-Chor1, and JHC7 were seeded in a white-walled 96-well plate in 100uL of media. Cells were treated in triplicate with a 3-fold serial dilution of two GCL inhibitors, Target 831-P2 and Target S, S-408-P2, with and without 1.5uM Ferrostatin-1, a ferroptosis inhibitor. UM-Chor1, U-CHCF365, and MUG-Chor1 cells were cultured in 4:1 IMDM/RPMI 1640 supplemented with 10% FBS, 1% NEAA, and 1% GlutaMAX. JHC7 cells were cultured in DMEM/F12 supplemented with 10% FBS. MUG-Chor1 cells were cultured on vessels coated with 50ug/mL collagen (Effect of collagen on chordoma cell growth). The top concentration of the GCL inhibitors was 10uM and the lowest concentration was 1.5nM. For the combination treatments, cells were plated in 1.5uM Fer-1. At the assay endpoint, 72 hours post-treatment, cell viability was measured using CellTiter-Glo 2.0 Cell Viability Assay. Experiments were repeated at least once to ensure reproducibility. Representative dose-response curves of the mean +/- the standard deviation are shown. The single agent data was normalized to the DMSO-only control. The combination treatment data was normalized to 1.5uM Fer-1.

The results indicate that chordoma cell lines do not respond to ferroptosis activation through GCL inhibition.

Further evaluation of ferroptosis in chordoma can be found here: Ferroptosis in chordoma