The antiestrogenic effects of black cohosh on BRCA1 and steroid receptors in breast cancer cells

Abstract

Background

Black cohosh (BC) is an herbal remedy often used by women to treat symptoms associated with menopause. Research has shown that the molecular activity of BC is associated with estrogen receptor alpha (ER-α) regulation. Progesterone receptor (PR) expression is found to be consistent with ER expression and mutations in the BRCA1 gene, a tumor-suppressor gene, are known to be responsible for about 40%–45% of hereditary breast cancers.

Purpose

The objective of this study was to determine the effects of BC alone, as well as in combination with hormones and antihormones, on cell viability and expression of ER-α, PR, and BRCA1 in both T-47D and MCF-7 cell lines.

Methods

Cells were cultured in charcoal-stripped serum prior to their treatment and subsequent protein extraction. Western blot analyses were performed following a Bio-Rad Bradford protein assay and SDS-PAGE gel electrophoresis, with ECL luminescence and Image Studio Lite software. Cellular viability assays were performed using propidium iodine (PI) staining, and the distribution of fluorescent structures was evaluated through confocal microscopy. RT-qPCR analysis was performed on extracted cellular RNA. All statistical analyses were performed using SPSS software, and data was subjected to Kruskal-Wallis testing, followed by post-hoc analysis using the Mann-Whitney U-test to determine the statistical significance of all findings.

Results

Western blot analysis displayed significant alterations of ER-α, PR, and BRCA1 protein levels after 24-hour treatment with 80–500 μM BC. BC displayed a concentration-dependent decrease on ER-α and BRCA1 expression, with an 87% reduction of ER-α expression and a 43% of BRCA1 expression in T-47D cells compared to control. After six days of treatment with 400 μM BC, a 50% decrease in cell proliferation was observed. Following 24 hours of co-treatment with 400 μM BC and 10 nM E₂, ER-α was downregulated by 90% and BRCA1 expression was reduced by 70% compared to control. The expression of PR, following the same treatment, exhibited similar effects. The proliferative effect of E₂ was reduced in the presence of BC.

Conclusion

Black Cohosh demonstrates substantial anti-cancer properties, and this study may significantly aid in the understanding of the molecular effects of BC on ER-α, PR, and BRCA1 in breast cancer cells.

Keywords:

• tumor suppressor
• MCF-7
• T-47D
• antitumor agent
• natural anticancer
• cancer treatment

Acknowledgments

We would like to thank Molly McDonald for her technical assistance. We would also like to thank NatureX for their donation of black cohosh. Funding was provided by the Center of Biomedical Research and Prevention Research Center, and by the Honors College at Oakland University, Rochester, MI, USA. An abstract of this paper was presented at the 98th Annual Meeting of the Endocrine Society in Boston April 1–4, 2016, as a poster presentation with interim findings. The poster’s abstract was published in “Poster Abstracts” in the meeting proceedings. Presentation Number: FRI-125: http://press.endocrine.org/doi/abs/10.1210/endo-meetings.2016. ED.1.FRI-125.

Disclosure

The authors report no conflicts of interest in this work.