Figure 4.
Etomoxir Inhibits Patient-derived GBM Cell Proliferation and Promotes Cell Death
(A) Quantification of the effects of 7-day 100 μM ETOM treatment administered twice per week across a panel of patient-derived GBM cell lines and two non-GBM controls (adult-mouse-derived neurospheres and human fetal-derived neurospheres) relative to vehicle-treated controls. n = 3 replicates, except for HK211 and HK207 in which only a single measurement was taken. Error bars = ± SEM. Significance values are relative to aSVZ NS with Bonferroni correction for multiple comparisons. Cultures that are EGFR amplified, EGFR amplified + EGFRviii mutant, IDH1 mutant, and PTEN deficient are indicated. See also Figure S3.
(B) Average 7-day growth of patient-derived cell line HK301with both CPT1A knockdown and 100 μM ETOM. n = 3 replicates. Univariate generalized linear model F(3,8) = 99.8, p = 1.12x10−6, post-hoc Tukey HSD relative to control (∗∗∗ = p < 0.001).
(C) Assessment of actively dividing cells by BrdU immunocytochemistry in two patient-derived GBM lines grown for 4 days in the presence of 100 μM ETOM. n = 9 replicates. Error bars = ± SEM. Scale bar, 30 μm.
(D) Flow cytometry analysis of cell death using dual Annexin V/PI staining in HK157 cells following 4-day treatment with 100 μM ETOM.
(E and F) (E) Effects of palmitic acid or (F) 3-OHB supplementation at different glucose concentrations on growth of HK157 cells in vitro. n = 3 replicates. Error bars = SD. Significance values represent t tests of each condition compared with the control for the same concentration of glucose (∗ = p < 0.05).
(G) Tumor growth measured by bioluminescence imaging for patient-derived cell line HK408. The effect of diet was not statistically significant by mixed effects model (p > 0.05). Numbers adjacent to lines indicate N at each time point.
(H) Kaplan-Meier curves for the data as in (E). Diet had no significant effect on survival by Mantel-Cox log rank values (p = 0.272).
See also Figure S4.