Fig. 7

Pimozide induces CML cell apoptosis by blocking STAT5A/miR-202-5p/USP15/Caspase-6 regulatory pathway. (a) K562 cells were treated with Pimozide (5 or 10 μM) for 48 h. Western blot analysis was used to detect protein levels of STAT5A and p-STAT5A. (b) K562 cells were prepared as (a), qRT-PCR was used to test miR-202-5p expression level. * P < 0.05, ** P < 0.01 vs DMSO control. (c) K562 cells were treated with Pimozide (5 μM) for 48 h. Western blot analysis was used to detect protein levels of USP15 and Caspase-6. (d) K562 cell were transfected with miR-202-5p mimic or mimic-NC and then treated with Pimozide (5 μM) or not. Western blot analysis was used to detect protein levels of USP15 and Caspase-6. (e) K562 cells were prepared as (c), Annexin V-FITC/PI staining was used to detect cell apoptosis. Right panel shows the apoptosis rate. ** P < 0.01 vs DMSO control. (f) K562G cells were treated with Imatinib (3 μM) and combination of Pimozide (5 μM) or not for 48 h. Annexin V-FITC/PI staining was used to detect cell apoptosis. Right panel shows the apoptosis rate. **P < 0.01 vs. DMSO control. (g) K562 cells were injected into the right posterior ankle of the nude mice to establish xenograft tumors. Mice were treated with Pimozide or not. Tumor volume was measured. ***P < 0.001 vs. control. (h) RNA was extracted from excised tumors and the miR-202-5p level was determined by qRT-PCR. *P < 0.05 vs. control. (i) Total proteins were extracted from excised tumors and the expression protein levels of STAT5, p-STAT5, USP15 and Caspase-6 were determined by Western blot analysis