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

Michael Crone,1,2 Kelly Hallman,1,2 Victoria Lloyd,1,2 Monica Szmyd,1,2 Briana Badamo,1,2 Mia Morse,1,2 Sumi Dinda1,2 1Department of Biomedical Diagnostic and Therapeutic Sciences, School of Health Sciences, Oakland University, Rochester, MI 48309-4476, USA; 2Institute for Stem Cell and Regenerative...

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Autores principales:	Crone M, Hallman K, Lloyd V, Szmyd M, Badamo B, Morse M, Dinda S
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2019
Materias:	Breast cancer tumor suppressor black cohosh MCF-7 T-47D anti-tumor agent Neoplasms. Tumors. Oncology. Including cancer and carcinogens RC254-282
Acceso en línea:	https://doaj.org/article/5c04d89295b140fa8ea16278ea0db0f9
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spelling	oai:doaj.org-article:5c04d89295b140fa8ea16278ea0db0f92021-12-02T01:27:39ZThe antiestrogenic effects of black cohosh on BRCA1 and steroid receptors in breast cancer cells1179-1314https://doaj.org/article/5c04d89295b140fa8ea16278ea0db0f92019-02-01T00:00:00Zhttps://www.dovepress.com/the-antiestrogenic-effects-of-black-cohosh-on-brca1-and-steroid-recept-peer-reviewed-article-BCTThttps://doaj.org/toc/1179-1314Michael Crone,1,2 Kelly Hallman,1,2 Victoria Lloyd,1,2 Monica Szmyd,1,2 Briana Badamo,1,2 Mia Morse,1,2 Sumi Dinda1,2 1Department of Biomedical Diagnostic and Therapeutic Sciences, School of Health Sciences, Oakland University, Rochester, MI 48309-4476, USA; 2Institute for Stem Cell and Regenerative Medicine and Center of Biomedical Sciences, Oakland University, Rochester, MI 48309-4476, USA 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 treatmentCrone MHallman KLloyd VSzmyd MBadamo BMorse MDinda SDove Medical PressarticleBreast cancertumor suppressorblack cohoshMCF-7T-47Danti-tumor agentNeoplasms. Tumors. Oncology. Including cancer and carcinogensRC254-282ENBreast Cancer: Targets and Therapy, Vol Volume 11, Pp 99-110 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	Breast cancer tumor suppressor black cohosh MCF-7 T-47D anti-tumor agent Neoplasms. Tumors. Oncology. Including cancer and carcinogens RC254-282
spellingShingle	Breast cancer tumor suppressor black cohosh MCF-7 T-47D anti-tumor agent Neoplasms. Tumors. Oncology. Including cancer and carcinogens RC254-282 Crone M Hallman K Lloyd V Szmyd M Badamo B Morse M Dinda S The antiestrogenic effects of black cohosh on BRCA1 and steroid receptors in breast cancer cells
description	Michael Crone,1,2 Kelly Hallman,1,2 Victoria Lloyd,1,2 Monica Szmyd,1,2 Briana Badamo,1,2 Mia Morse,1,2 Sumi Dinda1,2 1Department of Biomedical Diagnostic and Therapeutic Sciences, School of Health Sciences, Oakland University, Rochester, MI 48309-4476, USA; 2Institute for Stem Cell and Regenerative Medicine and Center of Biomedical Sciences, Oakland University, Rochester, MI 48309-4476, USA 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
format	article
author	Crone M Hallman K Lloyd V Szmyd M Badamo B Morse M Dinda S
author_facet	Crone M Hallman K Lloyd V Szmyd M Badamo B Morse M Dinda S
author_sort	Crone M
title	The antiestrogenic effects of black cohosh on BRCA1 and steroid receptors in breast cancer cells
title_short	The antiestrogenic effects of black cohosh on BRCA1 and steroid receptors in breast cancer cells
title_full	The antiestrogenic effects of black cohosh on BRCA1 and steroid receptors in breast cancer cells
title_fullStr	The antiestrogenic effects of black cohosh on BRCA1 and steroid receptors in breast cancer cells
title_full_unstemmed	The antiestrogenic effects of black cohosh on BRCA1 and steroid receptors in breast cancer cells
title_sort	antiestrogenic effects of black cohosh on brca1 and steroid receptors in breast cancer cells
publisher	Dove Medical Press
publishDate	2019
url	https://doaj.org/article/5c04d89295b140fa8ea16278ea0db0f9
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_version_	1718403074630877184

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

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