Mechanistic Evaluation of Black Cohosh Extract-Induced Genotoxicity in Human Cells

Seo, Ji-Eun; Guo, Xiaoqing; Petibone, Dayton M.; Shelton, Sharon D.; Chen, Ying; Li, Xilin; Tryndyak, Volodymyr; Smith-Roe, Stephanie L.; Witt, Kristine L.; Mei, Nan; Manjanatha, Mugimane G.

doi:10.1093/toxsci/kfab044

Mechanistic Evaluation of Black Cohosh Extract-Induced Genotoxicity in Human Cells

Journal Article · Thu Apr 15 00:00:00 EDT 2021 · Toxicological Sciences

DOI:https://doi.org/10.1093/toxsci/kfab044· OSTI ID:1808362

Seo, Ji-Eun ^[1]; ^[1]; Petibone, Dayton M. ^[1]; Shelton, Sharon D. ^[1]; Chen, Ying ^[1]; Li, Xilin ^[1]; Tryndyak, Volodymyr ^[2]; Smith-Roe, Stephanie L. ^[3]; Witt, Kristine L. ^[3]; ^[1]; ^[1]

Division of Genetic and Molecular Toxicology, National Center for Toxicological Research, Jefferson, Arkansas 72079, USA
Division of Biochemical Toxicology, National Center for Toxicological Research, Jefferson, Arkansas 72079, USA
Division of the National Toxicology Program, National Institute of Environmental Health Sciences (NIEHS), Research Triangle Park, North Carolina 27709, USA

Abstract

Black cohosh extract (BCE) is marketed to women as an alternative to hormone replacement therapy for alleviating menopausal symptoms. Previous studies by the National Toxicology Program revealed that BCE induced micronuclei (MN) and a nonregenerative macrocytic anemia in rats and mice, likely caused by disruption of the folate metabolism pathway. Additional work using TK6 cells showed that BCE induced aneugenicity by destabilizing microtubules. In the present study, BCE-induced MN were confirmed in TK6 and HepG2 cells. We then evaluated BCE-induced DNA damage using the comet assay at multiple time points (0.5–24 h). Following a 0.5-h exposure, BCE induced significant, concentration-dependent increases in %tail DNA in TK6 cells only. Although DNA damage decreased in TK6 cells over time, likely due to repair, small but statistically significant levels of DNA damage were observed after 2 and 4 h exposures to 250 µg/ml BCE. A G1/S arrest in TK6 cells exposed to 125 µg/ml BCE (24 h) was accompanied by apoptosis and increased expression of γH2A.X, p-Chk1, p-Chk2, p53, and p21. Conditioning TK6 cells to physiological levels of folic acid (120 nM) did not increase the sensitivity of cells to BCE-induced DNA damage. BCE did not alter global DNA methylation in TK6 and HepG2 cells cultured in standard medium. Our results suggest that BCE induces acute DNA strand breaks which are quickly repaired in TK6 cells, whereas DNA damage seen at 4 and 24 h may reflect apoptosis. The present study supports that BCE is genotoxic mainly by inducing MN with an aneugenic mode of action.

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Sponsoring Organization:: USDOE

OSTI ID:: 1808362

Journal Information:: Toxicological Sciences, Journal Name: Toxicological Sciences Journal Issue: 1 Vol. 182; ISSN 1096-6080

Publisher:: Oxford University PressCopyright Statement

Country of Publication:: United States

Language:: English

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