GM-CSF induces tumor cell EMT and stemness.

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GM-CSF induces tumor cell EMT and stemness. GM-CSF induces tumor cell EMT and stemness. A, IHC staining of UM5 tumor cells for the epithelial marker E-cadherin and mesenchymal marker vimentin following treatment with exogenous GM-CSF (100 ng/mL). B, Western blotting for EMT markers and phosphorylated (p) and total STAT3 in the UM5, UM2, and UM8 pancreatic cancer cells treated with exogenous GM-CSF. C, Western blot evaluating expression of EMT markers in either control or STAT3 siRNA (1 and 2) UM5 cells treated with GM-CSF. D, enhanced size of tumorspheres when UM5, UM2, and UM8 tumor cells were treated with 100 ng/mL GM-CSF (*, P < 0.0001 vs. control for UM5; *, P < 0.0001 vs. control for UM2; *, P < 0.004 vs. control for UM8). E, FACS analysis revealed an increase in the percentage of CSCs (ESA+CD44+CD24+) when cultured in the presence of exogenous GM-CSF (100 ng/mL; *, P < 0.05 vs. control for UM5; *, P < 0.02 vs. control for UM2; *, P < 0.0001 vs. control for UM8). F, FACS analysis for CSF2Ra receptor expression in CSCs (green) and bulk cells (red) showing significantly higher number of CSCs (green) expressing GM-CSF receptor than the bulk tumor cell population (red). Meghna Waghray et al. Cancer Discov 2016;6:886-899 ©2016 by American Association for Cancer Research