Page 131 - 2020_08-Haematologica-web

mTORi unlocks AML resistance to LSD1i
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Figure 1. In contrast to acute myeloid leukemia cells sensitive to LSD1 inhibition, mTOR signaling is robustly activated in resistant AML cells in response to their treatment with DDP38003. (A) Growth curves of the indicated acute myeloid leukemia (AML) cell lines treated with either vehicle or DDP38003 (0.5 mM) for the indicated time points of treatment assessed using trypan blue cell counting. Data were statistically analyzed using two way ANOVA followed by Bonferroni post-hoc test. A: P<0.05 compared to vehicle-treated cells (n=3). (B) Representative flowcytometry dot plots depicting the effect of six days of vehicle or DDP38003 (0.5 μM) treatment on the viability/apoptosis of the indicated AML cells assessed by Annexin V/PI staining (left panel) and their quantitation (right panel). (C) Western blot analysis of LSD1 levels in the indicated AML cell lines. b-actin served as the loading control. (D) Immunoblotting analysis of mTOR signaling pathway in LSD1i-sen- sitive (KASUMI-1, SKNO-1 and UF1) and LSD1i-resistant (NB4, OCI-AML3 and THP-1) AML cells treated for six days with either vehicle or different concentrations of DDP38003 (0.1 and 0.5 mM). Vinculin served as the loading control. The presented blots are derived from replicate samples run on parallel gels and controlled for even loading. LSD1: lysine specific demethylase-1; LSD1i: LSD1 inhibitors.
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