title: A Novel Combinatorial Epigenetic Therapy Using Resveratrol and Pterostilbene for Restoring Estrogen Receptor-α (ERα) Expression in ERα-Negative Breast Cancer Cells.
abstract: Breast cancer is the second most common cancer and a leading cause of cancer death in women. Specifically, estrogen receptor-α (ERα)-negative breast cancers are clinically more aggressive and normally do not respond to conventional hormone-directed therapies such as tamoxifen. Although epigenetic-based therapies such as 5-aza-2'-deoxycytidine and/or trichostatin A as DNA methyltransferase (DNMT) and histone deacetylase (HDAC) inhibitors, respectively, can regulate the expression of ERα, this can often lead to a number of side effects. Plant-based dietary compounds such as resveratrol and pterostilbene in novel combinatorial therapy provides new avenues to target these side effects and provide similar results with a higher level of safety. Here, we report that combinatorial resveratrol and pterostilbene leads to the reactivation of ERα expression in ERα-negative breast cancer cells in a time-dependent manner. Chromatin immunoprecipitation analysis of the ERα promoter in each cell type revealed an increase in enrichment of acetyl-H3, acetyl-H3lysine9 (H3K9) and acetyl-H4 active chromatin markers in the ERα promoter region after combinatorial treatment. This treatment also resulted in a significant change in HDAC and histone acetyl transferase (HAT) enzyme activity in these cells after 3 days of treatments. The combination resulted in a significant decrease in DNMT enzyme activity and 5-methylcytosine levels in MDA-MB-157 breast cancer cells. Moreover, reactivation of ERα expression by resveratrol combined with pterostilbene was found to sensitize ERα-dependent response to 17β-estradiol (E2)-mediated cellular proliferation and antagonist 4-hydroxytamoxifen (4-OHT)-mediated inhibition of cellular proliferation in ERα-negative breast cancer cells. E2 and 4-OHT further affected the ERα-responsive downstream progesterone receptor (PGR) gene in ERα reactivated MDA-MB-157 cells. Collectively, our findings provide a new and safer way of restoring ERα expression by regulating epigenetic mechanisms with the use of phytochemicals in combinatorial therapy. This combination can further provide effective treatment options for hormonal refractory breast cancer with available anti-hormonal therapy.
authors.affiliation: Department of Biology, University of Alabama Birmingham, 1300 University Boulevard, Birmingham, Alabama, 35294, United States of America.
authors.forename: Rishabh
authors.initials: R
authors.lastname: Kala
authors.affiliation: Department of Biology, University of Alabama Birmingham, 1300 University Boulevard, Birmingham, Alabama, 35294, United States of America.|Comprehensive Center for Healthy Aging, University of Alabama Birmingham, 1530 3rd Avenue South, Birmingham, Alabama, 35294, United States of America.|Comprehensive Cancer Center, University of Alabama Birmingham, 1802 6th Avenue South, Birmingham, Alabama, 35294, United States of America.|Nutrition Obesity Research Center, University of Alabama Birmingham, 1675 University Boulevard, Birmingham, Alabama, 35294, United States of America.|Comprehensive Diabetes Center, University of Alabama Birmingham, 1825 University Boulevard, Birmingham, Alabama, 35294, United States of America.
authors.forename: Trygve O
authors.initials: TO
authors.lastname: Tollefsbol
mesh_terms: D001943:Breast Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D019175:DNA Methylation; D044127:Epigenesis, Genetic; D047628:Estrogen Receptor alpha / Q000235:genetics / Q000378:metabolism*; D005260:Female; D006801:Humans; D011401:Promoter Regions, Genetic; D000077185:Resveratrol; D013267:Stilbenes / Q000627:therapeutic use*; D013629:Tamoxifen / Q000031:analogs & derivatives / Q000627:therapeutic use
chemical_list: D047628:Estrogen Receptor alpha; D013267:Stilbenes; D013629:Tamoxifen; C016601:afimoxifene; C107773:pterostilbene; D000077185:Resveratrol