1. In vivo AraC treatment does not induce any consistent changes in CD34+/−CD38+/− phenotypes nor in quiescent leukemic cells but increases apoptotic cell death in AraC-treated compared with PBS-treated AML-engrafted mice.
In vivo AraC treatment does not induce any consistent changes in CD34+/−CD38+/− phenotypes nor in quiescent leukemic cells but increases apoptotic cell death in AraC-treated compared with PBS-treated AML-engrafted mice. A and B, Flow cytometry analyses of human viable (Annexin V−/7-AAD−) CD45+CD33+ AML cells were performed to assess the percentage of CD34+CD38− cells in AraC-treated AML-xenografted mice compared with vehicle (PBS)-treated AML-xenografted mice (A) and stratified as following their basal level of CD34+CD38− phenotype (B). C and D, Hoechst/Pyronin Y–based flow cytometric assay was performed to measure cells in G0 in human viable (Annexin V−/7-AAD−) CD45+CD33+ AML cells in AraC-treated AML-xenografted mice compared with vehicle (PBS)-treated AML-xenografted mice. E, Representative histologic section showing Ki67 staining of the bone marrow. Scale bar, 100 μm. F, Ki67+ cycling cell quantification shows no difference between PBS-treated mice and AraC-treated mice. G, Expression level of genes involved in cell-cycle checkpoint and G0–G1 transition in residual AML cells after AraC treatment compared with PBS. a.u., arbitrary units. H and I, Percent of human apoptotic CD45+CD33+ AML cells was assessed by Annexin V/7-AAD–based flow cytometric assay in AraC-treated AML-xenografted mice compared with vehicle (PBS)-treated AML-xenografted mice. BM, bone marrow. Graphs of mean ± SEM. P values were determined by Mann–Whitney test (A, C, F, G, and H) or Wilcoxon matched-pairs signed rank test (B, D, and I). n.s., not significant; n.d., not determined. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤
Thomas Farge et al. Cancer Discov 2017;7:
©2017 by American Association for Cancer Research