ERK/MAPK regulates ERR[gamma] expression, transcriptional activity and receptor-mediated tamoxifen resistance in ER+ breast cancer

Selective estrogen receptor modulators such as tamoxifen (TAM) significantly improve breast cancer-specific survival for women with estrogen receptor-positive (ER+) disease. However, resistance to TAM remains a major clinical problem. The resistant phenotype is usually not driven by loss or mutation of the estrogen receptor; instead, changes in multiple proliferative and/or survival pathways over-ride the inhibitory effects of TAM. Estrogen-related receptor [gamma] (ERR[gamma]) is an orphan member of the nuclear receptor superfamily that promotes TAM resistance in ER+ breast cancer cells. This study sought to clarify the mechanism(s) by which this orphan nuclear receptor is regulated, and hence affects TAM resistance. mRNA and protein expression/phosphorylation were monitored by RT-PCR and western blotting, respectively. Site-directed mutagenesis was used to disrupt consensus extracellular signal-regulated kinase (ERK) target sites. Cell proliferation and cell-cycle progression were measured by flow cytometric methods. ERR[gamma] transcriptional activity was assessed by dual-luciferase promoter-reporter assays. We show that ERR[gamma] protein levels are affected by the activation state of ERK/mitogen-activated protein kinase, and mutation of consensus ERK target sites impairs ERR[gamma]-driven transcriptional activity and TAM resistance. These findings shed new light on the functional significance of ERR[gamma] in ER+ breast cancer, and are the first to demonstrate a role for kinase regulation of this orphan nuclear receptor.