Increased levels of EZH2, a critical regulator of cellular memory, signal the presence of
metastasis and poor outcome in
breast cancer patients. High levels of EZH2 are associated with nuclear pleomorphism, lack of
estrogen receptor expression, and decreased nuclear levels of BRCA1
tumor suppressor protein in invasive
breast carcinomas. The mechanism by which EZH2 overexpression promotes the growth of poorly differentiated invasive
carcinomas remains to be defined. Here, we show that EZH2 controls the intracellular localization of
BRCA1 protein. Conditional
doxycycline-induced upregulation of EZH2 in benign mammary epithelial cells results in nuclear export of
BRCA1 protein, aberrant mitoses with extra centrosomes, and
genomic instability. EZH2 inhibition in CAL51
breast cancer cells induces BRCA1 nuclear localization and rescues defects in ploidy and mitosis. Mechanistically, EZH2 overexpression is sufficient for activation of the
phosphoinositide 3-kinase/Akt (PI3K/Akt) pathway specifically through activation of Akt
isoform 1. EZH2-induced BRCA1 nuclear export,
aneuploidy, and mitotic defects were prevented by treatment with the PI3K inhibitors
LY294002 or
wortmannin. Targeted inhibition of Akt-1, Akt-2, and Akt-3
isoforms revealed that the EZH2-induced phenotype requires specific activation of Akt-1. The relevance of our studies to human
breast cancer is highlighted by the finding that high EZH2
protein levels are associated with upregulated expression of phospho-Akt-1 (Ser473) and decreased nuclear expression of phospho-BRCA1 (Ser1423) in 39% of invasive
breast carcinomas. These results enable us to pinpoint one mechanism by which EZH2 regulates BRCA1 expression and
genomic stability mediated by the PI3K/Akt-1 pathway.