Fig. 4

Effects of NCG on the viability, EST expression and estradiol metabolism in hepatocytes. A Evaluation of cytotoxicity in WRL-68 and Caco-2 cells for various NAR formulations (n = 3). B Assessment of the ability of multiple NAR formulations to activate EST genes at 24 h using the dual luciferase reporter gene assay (n = 3). Data are presented as mean ± SD, *p < 0.05, **p < 0.01, ***p < 0.001 indicates a comparison between the two groups. C Impact of NAR and NCG on estradiol metabolism in WRL-68 cells. D Influence of NAR and NCG on estrogen metabolism and E expression of EST enzyme in WRL-68 cells seeded at the lower chamber of a Transwell system after 24 h (n = 3). Data are presented as mean ± SD, ***p < 0.001, ^###p < 0.001 compared to the NAR group. Scar bar: 10 μm; F modulation of breast cancer growth and metastasis in a zebrafish xenograft model by NAR, NC, NG, and NCG through regulation of estradiol (40 μmol/L) metabolism. Statistical analysis of tumor fluorescence intensity and metastatic foci count in zebrafish (n = 6). Data are presented as mean ± SD, ns: p > 0.05, **p < 0.01, ***p < 0.001 compared to the E2 group; ^###p < 0.001 indicates a comparison between the two groups