The PI3K/AKT/mTOR signaling pathway is upregulated to mediate the increased aggressiveness in high AR–expressing AML cells. (A-B) Expression of (A) the PI3K gene (Pik3ca, Pik3cb, and Pik3cd) and (B) the AKT gene (Akt1 and Akt2) assessed by qPCR in low and high AR–expressing AML cells. The data were normalized to low AR–expressing AML cells and Gapdh housekeeping gene (n = 6-10 biologic replicates). (C) Western blot showing the expression of P-PI3K, PI3K, P-AKT, and AKT in low and high AR–expressing AML cells. Each lane represents 1 biologic replicate. (D) Western blot showing the expression of P-AKT and AKT in female and male patient–derived AML cells. Each lane represents 1 biologic replicate. (E) Western blot showing the expression of P-MTOR, mTOR, P-p70S6K, and p70S6K in low and high AR–expressing AML cells. Each lane represents 1 biologic replicate. (F) Flow cytometric analysis of apoptosis in high AR–expressing AML cells treated with 10 μM LY294002, 10 nM wortmannin, 10 μM Akt inhibitor, 10 nM sapanisertib, or 250 nM torin1 for 24 hours; apoptotic cells were identified as the Annexin V⁺7-AAD^–population. All data were normalized to and compared with vehicle treatment. (G) The survival curve of 2° male and female recipients transplanted with 1° Raptor TAM-iKO high AR–expressing AML cells. The mice were treated with tamoxifen (75 mg/kg per day in 100 μL corn oil, intraperitoneal injection for 5 days) at 1 week after transplantation and followed up for 60 days; ∗P < .05; ∗∗P < .01, as determined by the log-rank test. (H) Schematic illustration of the mechanism of action of finasteride and ARN509. Panels A-B, F, error bars represent the mean ± SEM of the replicates. ∗P < .05; ∗∗P < .01, as determined by a 1-tailed unpaired Student t test. TME, tumor microenvironment.