Fig. 6: Drug combination of 8GL and a mTORC1 inhibitor shows synergistic antileukemic effects in vitro and in vivo.
From: Activation of orphan receptor GPR132 induces cell differentiation in acute myeloid leukemia
A HL60 and MV4-11 cells were treated with vehicle, 30 μM 8GL, 3 μM everolimus (Eve), or drug combination for 72 h, and the expression of CD11b was determined using flow cytometry. Data are shown as mean ± SEM (n = 3). One-way ANOVA with Tukey’s multiple comparison tests was performed, *P < 0.05, **P < 0.01, ***P < 0.001. B Representative images of NBT reduction in HL60 and MV4-11 cells upon co-treatment of 8GL and Eve. Cells were treated as described in (A). Scale bars represent 20 μm. C Representative Wright–Giemsa staining images showing the mature myeloid cell morphology of HL60 and MV4-11 cells induced by 8GL and Eve combination. Cells were treated as described in (A). Scale bars represent 20 μm. D Representative images of colony formation of HL60 and MV4-11 cells incubated with 8GL (15 μM) and/or Eve (1.5 μM) for 7 days. The scale bar represents 100 μm. E Tumor growth curves of BALB/c nude mice inoculated with 1 × 106 HL60 cells. Mice were administrated with 8GL (30 mg/kg/d) in combination with Eve (3 mg/kg/d). Data are presented as mean ± SEM (n = 6). One-way ANOVA with Tukey’s multiple comparison tests was performed, *P < 0.05, **P < 0.01, ***P < 0.001. F Expression analysis of CD11b was determined using qPCR in the HL60 xenograft model as indicated in (E). Student’s t-tests were performed (n = 6), *P < 0.05, **P < 0.01, ***P < 0.001. G Bioluminescent imaging of NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ (NSG) mice transplanted with HL60-Luc cells treated with vehicle, 8GL (30 mg/kg), and/or everolimus (3 mg/kg). Representative images of two mice per group are shown. H Bioluminescence quantification representing tumor mass (count) on days post-injection of HL60-Luc cells. Mean normalized values are plotted for each treatment group. Data are presented as mean ± SEM (n = 6). Student’s t-tests were performed, *P < 0.05, **P < 0.01. I Kaplan–Meier overall survival plot of HL60-Luc xenograft AML mice model following treatment with vehicle, 8GL (30 mg/kg), and/or Eve (3 mg/kg) (n = 6). Statistical significance was tested using the log-rank test, *P < 0.05, **P < 0.01.
