Bioactive dietary supplements reactivate ER expression in ER-negative breast cancer cells by active chromatin modifications

Abstract

Breast cancer is the most common cancer and the leading cause of cancer death in women. Although tamoxifen therapy is successful for some patients, it does not provide adequate benefit for those who have estrogen receptor (ER)-negative cancers. Therefore, we approached novel treatment strategies by combining two potential bioactive dietary supplements for the reactivation of ERα expression for effective treatment of ER-negative breast cancer with tamoxifen. Bioactive dietary supplements such as green tea polyphenols (GTPs) and sulforaphane (SFN) inhibit DNA methyltransferases (DNMTs) and histone deacetylases (HDACs), respectively, which are of central importance to cancer prevention. In the present study, we have observed that treatment of ER-negative breast cancer cells with GTPs and SFN alone or in combination leads to the reactivation of ER expression. The combination of 20 µg/mL GTPs and 5 µM SFN was found to be the optimal dose of ER-reactivation at 3 days in MDA-MB-231 cells. The reactivation of ER expression was consistently correlated with ER promoter hypomethylation and hyperacetylation. Chromatin immunoprecipitation (ChIP) analysis of the ER promoter revealed that GTPs and SFN altered the binding of ER-transcriptional co-repressor complex thereby contributing to ERα-reactivation. In addition, treatment with tamoxifen in combination with GTPs and SFN significantly increased both cell death and inhibition of cellular proliferation in MDA-MB-231 cells in comparison to treatment with tamoxifen alone. Collectively, our findings suggest that a novel combination of bioactive-HDAC inhibitors with bioactive-demethylating agents is a promising strategy for the effective treatment of hormonal refractory breast cancer with available anti-estrogens.