Figure 7. AZD2014 is more effective than rapamycin or FRAX597 in decreasing cell viability in NF2-deficient primary meningioma cells.

A. Immunoblotting of a patient-derived, NF2-deficient primary meningioma cell line reveal attenuation of pS6K (mTORC1 readout) upon treatment with rapamycin (20 and 1000 nM, 1h; lanes 1 and 2) with no change in pAkt or pNDRG1 (mTORC2 readouts) compared to DMSO alone (lane 3). In contrast, treatment with AZD2014 (3-300 nM, 1h; lanes 4-8) shows dose-dependent inhibition of both mTORC1 activity (pS6K) and mTORC2 signaling (pAkt and pNDRG1). S6K, Akt, NDRG1 and GAPDH serve as controls. B and C. Dose-response curves for AZD2014 and rapamycin were determined for NF2-deficient primary (MN521, MN527, MN580) and immortalized (Ben-Men-1) meningioma cells lines, as well as primary (AC028) and immortalized arachnoid (AC007) cell lines, and dose-response curves for FRAX597 were carried out in NF2-deficient primary (MN320, MN521, MN548) and Ben-Men-1 meningioma cells lines as indicated. Cells were exposed to increasing concentrations of rapamycin (B, left panel) in a 10 point, 10-fold serial dilution series (0 - 25 μM); AZD2014 (B, right panel) in a 10 point, 4-fold serial dilution series (0 - 20μM); or FRAX597 C. in a 10 point, 5-fold serial dilution series (0 - 25 μM) for 72h. Cell viabilities were assessed using CellTiter-Glo assays and plotted as % relative to DMSO controls. Cell viability and IC₅₀ measurements were performed in at least three independent experiments with similar results. Data are presented as mean +/− SD for 3 replicates/drug dosage point B., or 4 replicates/drug dosage point C..