Fig. 7

NFIL3 promotes the proliferation, migration and invasion of TNBC cells by activating the NF-κB inflammatory pathway through inhibiting NFKBIA. A. The restored expression of NFKBIA in Hs578T cells with NFIL3 overexpression was confirmed by western blot. The relative NFKBIA expression was analyzed using Image J software (***p < 0.001). B. Translocation of NF-κB detected using an immunofluorescence assay was employed to investigate the effect of NFKBIA restoration on the activation of NF-κB signaling in Hs578T cells with NFIL3 overexpression. (magnification, 400×, scale bars = 50 μm.) C. A luciferase reporter assay was used to detect the effect of NFKBIA restoration on the activity of NF-κB signaling in Hs578T cells upon NFIL3 overexpression with or without TNF-α (25 ng/mL) stimulation (**p < 0.01, ***p < 0.001). The experiment was repeated in triplicate. D. The effect of NFKBIA restoration on the proliferation of Hs578T cells with NFIL3 overexpression (*p < 0.05, **p < 0.01, ***p < 0.001). E-F. The effects of NFKBIA restoration on the migration and invasion of Hs578T cells with NFIL3 overexpression (**p < 0.01). G. Aspirin (4 mmol/L) and Bay11–7082 (10 μmol/L) blocked the activation of NF-κB signaling triggered by NFIL3 overexpression or TNF-α (25 ng/mL) stimulation (*p < 0.05, **p < 0.01, ***p < 0.001). H. Both aspirin (4 mmol/L) and Bay11–7082 (10 μmol/L) significantly inhibited the promoting effect of NFIL3 on the proliferation of Hs578T cells (***p < 0.001). I-J. Both aspirin (4 mmol/L) and Bay11–7082 (10 μmol/L) significantly inhibited the promotional effect of NFIL3 on the migration and invasion of Hs578T cells (**p < 0.01, ***p < 0.001). K. Aspirin suppressed the promotional roles of NFIL3 in promoting the growth of Hs578T cells in a xenograft mouse model by evaluating the weight and volume of tumors (*p < 0.05, **p < 0.01). L. Aspirin suppressed the promotional roles of NFIL3 in promoting the lung metastasis of Hs578T cells using a tail vein injection mouse model (magnification, 100×; scale bars = 200 μm; the black arrow indicates the metastatic foci; *p < 0.05)