Figure 6.
Pharmacological inhibition of PRL2 reduces leukemia burden and extends the survival of mice transplanted with primary human AML cells. (A) PRLi treatment decreased the viability of primary human AML cells with FLT3-ITD mutation in a dosage-dependent manner. (B) PRLi treatment reduced the colony forming ability of primary human AML cells with or without FLT3-ITD mutation. Samples 3153 and 3202 are from AML patients with WT FLT3, whereas samples 3142 and 3179 are from AML patients with FLT3-ITD. (C) Cell cycle analysis of primary AML cells with FLT3-ITD mutation (AML3242) at 24 hours following DMSO or PRLi (10 μM) treatment. (D) Apoptosis analysis of primary AML cells with FLT3-ITD (AML3242) at 24 hours following DMSO or PRLi (10 μM) treatment. (E) Kaplan-Meier survival curve of NSG mice transplanted with 4 × 106 human CD45+ leukemia cells (AML3179) following 3 weeks of DMSO or PRLi treatment (n = 6 mice per group). (F) Flow cytometry analysis of human CD45+ cells in PB, BM, and spleen of NSG mice transplanted with 4 × 106 human CD45+ leukemia cells (AML3179) after 3 weeks of DMSO or PRLi treatment (n = 4 mice per group). (G) Representative western blot analysis of AKT, STAT5, and ERK phosphorylation in Prl2+/+, Prl2−/−, Flt3+/ITD, Flt3+/ITDPrl2−/−, Flt3ITD/ITD, and Flt3ITD/ITDPrl2−/− BM mononuclear cells. (H) Representative western blot analysis of FLT3, AKT, STAT5, and ERK phosphorylation in MV-4-11 cells expressing shCtrl, shPRL2, or shPRL2#2 (left) and following 24 hours of DMSO or 5 μM PRLi treatment (right). (I) Representative western blot analysis of FLT3, AKT, STAT5, and ERK phosphorylation in primary AML cells with FLT3-ITD mutation (AML3080 and AML3220) following 24 hours of DMSO or PRLi (10 μM) treatment. (J) Representative western blot analysis of AKT, STAT5, and ERK phosphorylation in human CD45+ cells isolated from the BM of NSG mice at 4 weeks after transplantation with MV-4-11 cells expressing shCtrl or shPRL2 (left panel, n = 3 mice per group); human CD45+ cells in the BM of NSG mice transplanted with MV-4-11 cells following 3 weeks of DMSO or PRLi treatment (right panel, n = 3 mice per group). (K) Representative western blot analysis of AKT, STAT5, and ERK phosphorylation in human CD45+ cells isolated from the BM of NSG mice transplanted with patient-derived xenograft cells (AML3179) following 3 weeks of DMSO or PRLi treatment (n = 3 mice per group).

Pharmacological inhibition of PRL2 reduces leukemia burden and extends the survival of mice transplanted with primary human AML cells. (A) PRLi treatment decreased the viability of primary human AML cells with FLT3-ITD mutation in a dosage-dependent manner. (B) PRLi treatment reduced the colony forming ability of primary human AML cells with or without FLT3-ITD mutation. Samples 3153 and 3202 are from AML patients with WT FLT3, whereas samples 3142 and 3179 are from AML patients with FLT3-ITD. (C) Cell cycle analysis of primary AML cells with FLT3-ITD mutation (AML3242) at 24 hours following DMSO or PRLi (10 μM) treatment. (D) Apoptosis analysis of primary AML cells with FLT3-ITD (AML3242) at 24 hours following DMSO or PRLi (10 μM) treatment. (E) Kaplan-Meier survival curve of NSG mice transplanted with 4 × 106 human CD45+ leukemia cells (AML3179) following 3 weeks of DMSO or PRLi treatment (n = 6 mice per group). (F) Flow cytometry analysis of human CD45+ cells in PB, BM, and spleen of NSG mice transplanted with 4 × 106 human CD45+ leukemia cells (AML3179) after 3 weeks of DMSO or PRLi treatment (n = 4 mice per group). (G) Representative western blot analysis of AKT, STAT5, and ERK phosphorylation in Prl2+/+, Prl2−/−, Flt3+/ITD, Flt3+/ITDPrl2−/−, Flt3ITD/ITD, and Flt3ITD/ITDPrl2−/− BM mononuclear cells. (H) Representative western blot analysis of FLT3, AKT, STAT5, and ERK phosphorylation in MV-4-11 cells expressing shCtrl, shPRL2, or shPRL2#2 (left) and following 24 hours of DMSO or 5 μM PRLi treatment (right). (I) Representative western blot analysis of FLT3, AKT, STAT5, and ERK phosphorylation in primary AML cells with FLT3-ITD mutation (AML3080 and AML3220) following 24 hours of DMSO or PRLi (10 μM) treatment. (J) Representative western blot analysis of AKT, STAT5, and ERK phosphorylation in human CD45+ cells isolated from the BM of NSG mice at 4 weeks after transplantation with MV-4-11 cells expressing shCtrl or shPRL2 (left panel, n = 3 mice per group); human CD45+ cells in the BM of NSG mice transplanted with MV-4-11 cells following 3 weeks of DMSO or PRLi treatment (right panel, n = 3 mice per group). (K) Representative western blot analysis of AKT, STAT5, and ERK phosphorylation in human CD45+ cells isolated from the BM of NSG mice transplanted with patient-derived xenograft cells (AML3179) following 3 weeks of DMSO or PRLi treatment (n = 3 mice per group).

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal