Research Paper Volume 13, Issue 10 pp 13644—13662

microRNA-1246-containing extracellular vesicles from acute myeloid leukemia cells promote the survival of leukemia stem cells via the LRIG1-meditated STAT3 pathway

AML cell-derived EVs elevated the viability and colony formation and reduced the apoptosis and differentiation of LSCs. (A) Absorption of CFSE-labeled AML cell-derived EVs by LSCs observed under a fluorescence microscope (CFSE: green, DAPI: blue; × 500). (B) Fluorescence microscopic observation of Cy3 labeled AML cell-derived EVs co-cultured with LSCs (Cy3-miR-1246: red, DAPI: blue, FITC-LSCs: green; × 400). (C) Viability of LSCs after exposure to AML cell-derived EVs evaluated by MTS assay. (D) Effects of AML cell-derived EVs on the colony formation capacity of LSCs identified by colony formation assay. (E) Effects of AML cell-derived EVs on LSCs apoptosis assessed by flow cytometry. (F) Changes in differentiation of LSCs after treatment with AML cell-derived EVs detected by differentiation assay. * p vs. LSCs treated with PBS. Data in the figures are all measurement data, expressed as mean ± standard deviation. Independent sample t-test was applied for comparison between two groups. Data among multiple groups were compared by one-way ANOVA. The experiments were repeated in triplicate.

Figure 2. AML cell-derived EVs elevated the viability and colony formation and reduced the apoptosis and differentiation of LSCs. (A) Absorption of CFSE-labeled AML cell-derived EVs by LSCs observed under a fluorescence microscope (CFSE: green, DAPI: blue; × 500). (B) Fluorescence microscopic observation of Cy3 labeled AML cell-derived EVs co-cultured with LSCs (Cy3-miR-1246: red, DAPI: blue, FITC-LSCs: green; × 400). (C) Viability of LSCs after exposure to AML cell-derived EVs evaluated by MTS assay. (D) Effects of AML cell-derived EVs on the colony formation capacity of LSCs identified by colony formation assay. (E) Effects of AML cell-derived EVs on LSCs apoptosis assessed by flow cytometry. (F) Changes in differentiation of LSCs after treatment with AML cell-derived EVs detected by differentiation assay. * p < 0.05 vs. LSCs treated with PBS. Data in the figures are all measurement data, expressed as mean ± standard deviation. Independent sample t-test was applied for comparison between two groups. Data among multiple groups were compared by one-way ANOVA. The experiments were repeated in triplicate.