Fig. 2

MiR-141 reprograms stromal fibroblasts to be oncogenic drivers, and GROα is a major proinflammatory cytokine (A) The XTT cell proliferation assay indicated that stromal cell-conditioned medium (SCCM) from miR-141-transfected WPMY-1 cells (SCCM miR-141) increased the cell proliferation of ES-2 and SKOV3 cells on Day 2 and Day 3, compared with scrambled control medium from WPMY-1 cells (SCCM Ctrl) (n = 5, Mean ± SD, t-test, *P < 0.05, **P < 0.01). N = 1 independent experiment. (B) Transwell cell migration/invasion assays demonstrated that SCCM miR-141 treatment had convincingly induced a higher capacity of migration at 16 h or invasion at 24 h in ES-2 and SKOV3 cells as compared with SCCM Ctrl treatment (n = 6, mean ± SEM, t test, **P < 0.01, ***P < 0.001). Scale bar =100 μm. (C) Human XL cytokine array analysis revealed the number of inflammatory cytokines such as GROα in the stromal cell-conditioned medium derived from WPMY-1 scrambled control cells (SCCM Ctrl) and miR-141 overexpressing cells (SCCM miR-141). N = 2 independent experiments. (D) The XTT cell proliferation assay showed a dose-dependent increase in cell proliferation of ES-2 and OVCA433 cells upon a 4-day incubation with recombinant GROα (50 ng/mL) as compared with the respective untreated control (n = 6, mean ± SEM, two-way ANOVA, **P < 0.01, ***P < 0.001, ****P < 0.0001). N = 3 independent experiments. (E) Transwell cell migration/invasion assays showed that the recombinant GROα (60 ng/mL) remarkably promoted cell migration/invasion capacity in ES-2 and OVCA433 cells after 14 h (migration) and 20 h (invasion) as compared with the respective untreated control (Ctrl) (n = 6, mean ± SEM, t-test, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001). Scale bar = 100 μm