Figure 7.

Treatment with EMs induces apoptosis, loss of cell viability of PD somatic and germline mtRUNX1 expressing AML, depletes RUNX1 protein expression, and improves the survival of mice engrafted with mtRUNX1 expressing AML cells. (A) OCI-AML5 cells were treated with anisomycin, cinobufagin, CD1530, narciclasine, fenbendazole, or mevastatin at concentrations ranging from 10 nM to 20 µM for 48 hours. The IC₅₀ dose of each drug was calculated and is shown. (B-C) Immunoblot analysis of OCI-AML5 cells treated with the indicated concentrations of EMs for 24 hours. Expression levels of β-actin in the lysates served as the loading control. (D) Kaplan-Meier survival plot of the in vivo activity of 1 mg/kg of narciclasine, 50 mg/kg or 100 mg/kg of fenbendazole, or 1 mg/kg of cinobufagin (twice daily) against OCI-AML5/GFP-Luc xenografts in NSG mice. Significance between mice treated with 1 mg/kg of narciclasine, 50 mg/kg or 100 mg/kg of fenbendazole, or 1 mg/kg of cinobufagin (twice daily) vs vehicle-treated mice was determined by using a Mantel-Cox rank sum test. (E-G) PD CD34+ somatic mtRUNX1 expressing AML cells (n = ≥6 samples per drug), wtRUNX1 AML cells (n = 6), germline mtRUNX1 expressing AML cells (n = ≥3), and mtRUNX1 FPD cells (n = ≥3) were treated with the indicated concentrations of anisomycin, cinobufagin, and narciclasine for 48 hours. The percentage of propidium iodide (PI)–positive nonviable cells was then determined by using flow cytometry. (H) Normal CD34⁺ cord blood progenitor cells (n = 3) were treated with the indicated concentrations of anisomycin, cinobufagin, and narciclasine for 48 hours. The percentage of PI-positive nonviable cells was then determined by using flow cytometry.