Fig 4. Glycolysis inhibition with clotrimazole affects glioma cell survival in vitro and delays tumor growth in vivo.

A. The IC₅₀ of different glycolysis inhibitors was determined for patient derived GBM cells (P3A) and normal human astrocytes (NHA). N: normoxia, H: hypoxia (0.1% O2). Cells were exposed to indicated compounds for 72h and IC₅₀ was determined with the SRB assay (n = 3). B. The cytotoxic effect of clotrimazole (30μM) was assessed on organotypic spheroids of several patient-derived GBM (P3, P8, T16) and NHA, treated for 72h in normoxia and 0,1% O₂ (n = 5). Representative images showing viable cells in ‘green’, dead cells in ‘red’ fluorescence. C. P3 spheroids were implanted intracranially and clotrimazole (CTZ, 150mg/kg) treatment was started 3 weeks after implantation (n = 7). Kaplan-Meier curve shows significantly improved mouse survival (* p<0.05).