Fig. 10

The anti-CSC effects of ropivacaine on MCF-7 cells is dependent on NF-κB/GGT1 feedback loop. MCF-7 cells was transfected with GGT1 overexpression plasmid (GGT1-OE) or empty plasmid (Vector) and treated with ropivacaine (10 μM) or negative control for 48 h. A RNA level of GGT1 in the indicated treated-MCF-7 cells was determined by qRT-PCR. B Protein levels of GGT1, p-iκBa, iκBa, p-NF-κB and NF-κB in the indicated treated-MCF-7 cells were examined by western blot. C Stem cell markers (CD133, OCT4, SOX2) in the indicated treated-MCF-7 cells were analyzed by western blot. D CD44⁺/CD24.⁻ subpopulation in the indicated treated-MCF-7 cells was measured by FACS analysis. E Mammosphere formation of the indicated treated-MCF-7 cells (scale bar = 100 μm). F Migration and invasion of the indicated treated-MCF-7 cells were examined by transwell assays (scale bar = 100 μm). G Cell viability of MCF-7/ADR cells transfected with GGT1 overexpression plasmid (GGT1-OE) or empty plasmid (Vector) and treated with ropivacaine (10 μM) or negative control for 48 h as indicated, exposure to the indicated concentration of ADR. H Cell apoptosis of the indicated treated-MCF-7/ADR cells exposed to ADR (10 μM) were assessed by FACS analysis. Results are shown are shown as mean ± S.D from at least three independent experiments.. *p < 0.05; **p < 0.01; ***p < 0.001 (Two-way ANOVA test in G, others unpaired two-tailed Student’s t test)