Figure 5. USP7/PHF8/cyclin A2 signaling pathway promotes breast carcinogenesis.

(A) IHC analysis of the expression levels of USP7, PHF8, and cyclin A2 in breast tumors. Representative images (original magnification, ×200) from normal mammary tissue and breast carcinoma samples at different stages are shown (top panel). The values of the stainings were determined by Image-Pro Plus software and are presented with box plots (middle panel). *P < 0.05, **P < 0.01, 1-way ANOVA. The correlation coefficient and P values were analyzed as indicated (bottom panel). (B) Western blotting analysis of the protein levels of USP7, PHF8, and cyclin A2 in human mammary epithelial cell (HMEC) and breast cancer cell lines (left panel), or in paired samples from infiltrating ductal breast carcinomas and adjacent breast tissues (right panel). (C) MCF-7 tumors stably expressing shRNAs and genes as indicated were transplanted onto athymic mice (n = 6), and tumor volumes were measured weekly (top panel). Representative tumors and sacrificed mice are shown (middle panel). The expression levels of the indicated proteins in these tumors were examined by Western blotting (bottom panel). Each bar represents the mean ± SD. *P < 0.05 and **P < 0.01, 2-way ANOVA.