Figure 1 from Cyclin-Dependent Kinase-9 and Oxidative Phosphorylation Inhibition Overcomes Ibrutinib Resistance in Mantle Cell Lymphoma

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AZD4573 induces apoptosis and thwarts survival of MCL cells. A, MCL cell lines were treated with the indicated doses of AZD4573 for 72 hours. Cell proliferation was assessed using a colorimetric tetrazolium-based assay. The mean ± SEM is shown. IC50 value was calculated using GraphPad Prism software set to variable slope. B, Cells were treated with indicated doses of AZD4573 for 8 hours. Apoptosis was determined using Annexin V–FITC staining. Data are the mean ± SEM. *, P < 0.05 and **, P < 0.01 vs. untreated control. C, Primary MCL cells (n = 5 individual samples tested in technical triplicates) were cocultured with stromal cells for 24 hours and then treated with indicated doses of AZD4573 for 24 hours. Apoptosis of the CD19+ B-cell population was quantified using Annexin V–FITC staining. *, P < 0.05 and **, P < 0.01 vs. untreated control. D, Parental and IR MCL cell lines were treated with 10 nmol/L of AZD4573 or vehicle control for the indicated time points. Cell lysates were subjected to immunoblotting. E, Mice were inoculated with MCL. Two distinct MCL PDX models were used. Once circulating CD5+/CD19+ MCL cells were detected in the peripheral blood, mice began treatment as indicated in the methods (n = 3–11 per group). Kaplan–Meier survival curves are shown, with significance determined by the log-rank test. *, P < 0.05; **, P < 0.01.

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