Abstract
The purpose of this study was to determine whether black cohosh contains constituents that inhibit the growth of human breast cancer cells, and therefore might eventually be useful in the prevention or treatment of breast cancer. Black cohosh rhizomes were extracted with methanol/water and fractionated by solvent–solvent partitioning to yield three fractions: hexane, ethyl acetate and water. The ethyl acetate fraction displayed the highest potency in two cell-based assays, growth inhibition and cell cycle analysis. This fraction inhibited growth of both the ER+ MCF7 and ER−MDA-MB-453 human breast cancer cell lines with IC50 values of about 20 and 10 µg/ml, respectively. It also induced cell cycle arrest at G1 when tested at 30 µg/ml and at G2/M at 60 µg/ml in MCF7 cells. This suggests that the extract contains a mixture of components with the more active (or more abundant) causing G1 arrest and the less active causing G2/M arrest. We then examined specific components in this extract. The triterpene glycoside fraction obtained by polyamide column chromatography, and the specific triterpene glycosides actein, 23-epi-26-deoxyactein and cimiracemoside A, inhibited growth of the MCF7 human breast cancer cells and induced cell cycle arrest at G1. The most potent compound, actein, decreased the level of cyclin D1, cdk4 and the hyperphosphorylated form of the pRb protein and increased the level of p21cip1 in MCF7 cells, changes that may contribute to the arrest in G1. Further studies are in progress to identify the mechanisms by which actein and related compounds present in black cohosh inhibit growth of human breast cancer cells.
Similar content being viewed by others
References
Sporn MB, Suh N: Chemoprevention of Cancer. Carcinogenesis 21: 525–530, 2000
McKenna DJ, Jones K, Humphrey S, Hughes K: Black cohosh: efficacy, safety, and use in clinical and preclinical applications. Altern Ther 7: 93–100, 2001
Foster S: Black cohosh: Cimicifuga racemosa. A literature review. HerbalGram 45: 35–49, 1999
Hsu H-Y, Chen Y-P, Hsu C-S, Shen S-J, Chen C-C, Chang H-C: Oriental Materia Medica: A Concise Guide. Keats Publishing, Inc., New Canaan, 1986
Upton R: Black cohosh rhizome. In: American Herbal Pharamacopoeia and Therapeutic Compendium. American Herbal Pharmacopoeia, Santa Cruz, 2002
Lehmann-Willenbrock E, Riedel HH: Clinical and endocrinological examinations concerning therapy of climacteric symptoms following hysterectomy with remaining ovaries. Zent Bl Gynakol 110: 611–618, 1998
Stoll W: Phytotherapy influences atrophic vaginal epithelium: Double-blind study-Cimicifuga vs. estrogenic substances. Therapeuticum 1: 23–31, 1997
Chen SN, Li W, Fabricant DS, Santarsiero BD, Mesecar A, Fitzloff JF, Fong HH, Farnsworth NR: Isolation, structure elucidation, and absolute configuration of 26-deoxyactein from Cimicifuga racemosa and clarification of nomenclature associated with 27-deoxyactein. J Natl Prod 65: 601–605, 2000
Zheng QY, He K, Pilkington L, Yu S, Zheng B: CimiPure (Cimicufuga racemosa): a standardized black cohosh extract with novel triterpene glycoside for menopausal women. In: Shahidi F, Ho C-T (eds) Phytochem Phytopharm. AOCS Press, Champagne, 2000, pp 360–370
Loser B, Kruse SO, Melzig MF, Nahrstedt A: Inhibition of neutrophil elastase activity by cinnamic acid derivatives from Cimicifuga racemosa. Planta Med 66: 751–753, 2000
Burdette JE, Chen SN, Lu ZZ, Xu H, White BE, Fabricant DS, Liu J, Fong HH, Farnsworth NR, Constantinou AI, Van Breemen RB, Pezzuto JM, Bolton JL: Black cohosh (Cimicifuga racemosa L.) protects against menadione-induced DNA damage through scavenging of reactive oxygen species: bioassay-directed isolation and characterization of active principles. J Agric Food Chem 50: 7022–7028, 2002
Dixon-Shanies D, Shaikh N: Growth inhibition of human breast cancer cells by herbs and phytoestrogens. Oncol Rep 6: 1383–1387, 1999
Nesselhut T, Schellhase C, Dietrich R, Kuhn W: Studies on mammary carcinoma cells regarding the proliferative potential of herbal medications with estrogen-like effects. Arch Gynecol Obstet 254: 817–818, 1993
Bodinet C, Freudenstein J: Influence of Cimicifuga racemosa on the proliferation of estrogen receptor-positive human breast cancer cells. Breast Cancer Res Treat 76: 1–10, 2002
Watanbe K, Mimaki Y, Sakagami H, Sashida Y: Cycloartane glycosides from the rhizomes of Cimicifuga racemosa and their cytotoxic activities. Chem Pharm Bull (Tokyo) 50: 121–125, 2002
Kennelly EJ, Gerhauser C, Song LL, Graham JG, Beecher CWW, Pezzuto JM, Kinghorn AD: Induction of quinone reductase by withanolides isolated from Physalis philadelphica (tomatillos). J Agric Food Chem 45: 3771–3777, 1997
Lim JTE, Piazza GA, Han K-H, Delohery TM, Li H, Finn TS, Buttyan R, Yamamoto H, Sperl GJ, Brendel K, Gross PH, Pamukcu R, Weinstein IB: Sulindac derivatives inhibit growth and induce apoptosis in human prostate cancer cell lines. Biochem Pharmacol 58: 1097–1107, 1999
Luo J, Soh J-W, Xing W-Q, Mao Y, Matsuno T, Weinstein IB: PM-3, a benzo-g-pyran derivative isolated from propolis, inhibits growth of MCF-7 human breast cancer cells. Anticancer Res 21: 1665–1672, 2001
Han EK, Sgambato A, Jiang W, Zhang YJ, Santella RM, Doki Y, Cacace AM, Schieren I: Stable overexpression of cyclin D1 in a human mammary epithelial cell line prolongs the S-phase and inhibits growth. Oncogene 10: 953–961, 1995
Sgambato A, Zhang Y-J, Ciaparrone M, Soh J-W, Cittadini A, Santella RM, Weinstein IB: Overexpression of p27(Kip1) inhibits the growth of both normal and transformed human mammary epithelial cells. Cancer Res 58: 3448–3454, 1998
Kruse SO, Löhning A, Pauli GF, Winterhoff H, Nahrstedt A: Fukiic and piscidic acid esters from the rhizome of Cimicifuga racemosa and the in vitro estrogenic activity of fukinolic acid. Planta Med 65: 763–764, 1999
Tsutsui S, Ohno S, Murakami S, Hachitanda Y, Oda S: Prognostic value of c-erbB2 expression in breast cancer. J Surg Oncol 79: 216–233, 2002
Weinstein IB: Disorders of cell circuitry during multistage carcinogenesis: the role of homeostasis. Carcinogenesis 21: 857–864, 2000
Joe AK, Arber N, Bose S, Heitjan D, Zhang Y, Weinstein IB, Hibshoosh H: Cyclin D1 overexpression is more prevalent in non-Caucasian breast cancer. Anticancer Res 21: 3535–3539, 2001
Han EK, Arber N, Yamamoto H, Lim JT, Delohery T, Pamukcu R, Piazza GA, Xing WQ, Weinstein IB: Effects of sulindac and its metabolites on growth and apoptosis in human mammary epithelial and breast carcinoma cell lines. Breast Cancer Res Treat 48: 195–203, 1998
Masuda M, Suzui M, Weinstein IB: Effects of epigallocatechin-3-gallate on growth, epidermal growth factor receptor signaling pathways, gene expression, and chemosensitivity in human head and neck squamous cell carcinoma cell lines. Clin Cancer Res: 4220–4229, 2001
Meiers S, Kemeny M, Weyand U, Gastpar R, von Angerer E, Marko D: The anthocyanidins cyanidin and delphinidin are potent inhibitors of the epidermal growth-factor receptor. J Agric Food Chem 49: 958–962, 2001
Haridas V, Higuchi M, Jayatilake GS, Bailey D, Mujoo K, Blake ME, Arntzen CJ, Gutterman JU: Avicins: triterpenoid saponins from Acacia victoriae (Bentham) induce apoptosis by mitochondrial perturbation. Proc Natl Acad Sci USA 98: 5821–5826, 2001
Pisha E, Chai H, Lee IS, Chagwedera TE, Farnsworth NR, Cordell GA, Beecher CW, Fong HH, Kinghorn AD, Brown DM et al.: Discovery of betulinic acid as a selective inhibitor of human melanoma that functions by induction of apoptosis. Nat Med 1: 1046–1051, 1995
Fulda S, Debatin KM: Betulinic acid induces apoptosis through a direct effect on mitochondria in neuroectodermal tumors. Med Pediatr Oncol 35: 616–618, 2000
Suh N, Honda T, Finlay H, Barchowsky A, Williams C, Benoit N, Xie OW, Nathan C, Gribble G, Sporn M: Novel triterpenoids suppress inducible nitric oxide synthase (iNOS) and inducible cyclooxygenase (COX-2) in mouse macrophages. Cancer Res 58: 717–723, 1998
Suh N, Wang Y, Honda T, Gribble GW, Dmitrovsky E, Hickey WF, Maue RA, Place AE, Porter DM, Spinella MJ, Williams CR, Wu G, Dannenberg AJ, Flanders KC, Letterio JJ, Mangelsdorf DJ, Nathan CF, Nguyen L, Porter WW, Ren RF, Roberts AB, Roche NS, Subbaramaiah K, Sporn MB: A novel synthetic oleanane triterpenoid, 2-cyano-3,12-dioxoolean-1,9-dien-28-oic acid, with potent differentiating, antiproliferative, and anti-inflammatory activity. Cancer Res 59: 336–341, 1999
Ito Y, Pandey P, Sporn MB, Datta R, Kharbanda S, Kufe D: The novel triterpenoid CDDO induces apoptosis and differentiation of human osteosarcoma cells by a caspase-8 dependent mechanism. Mol Pharmacol 5: 1094–1099, 2001
Pedersen IM, Kitada S, Schimmer A, Kim Y, Zapata JM, Charboneau L, Rassenti L, Andreeff M, Bennett F, Sporn MB, Liotta LD, Kipps TJ, Reed JC: The triterpenoid CDDO induces apoptosis in refractory CLL B cells. Blood 8: 2965–2972, 2002
Stadheim TA, Suh N, Ganju N, Sporn MB, Eastman A: The novel triterpenoid 2-cyano-3,12-dioxooleana-1,9-dien-28-oic acid (CDDO) potently enhances apoptosis induced by tumor necrosis factor in human leukemia cells. J Biol Chem 19: 16448–16455, 2002
Wang Y, Porter WW, Suh N, Honda T, Gribble GW, Leesnitzer LM, Plunket KD, Mangelsdorf DJ, Blanchard SG, Willson TM, Sporn MB: A synthetic triterpenoid, 2-cyano-2,12-dioxooleana-1,9-dien-28-oic acid (CDDO), is a ligand for the peroxisome proliferator-activated receptor gamma. Mol Endocrinol 14: 1550–1556, 2000
Liske E: Therapeutic efficacy and safety of Cimicifuga racemosa for gynecologic disorders. Adv Nat Ther 15: 45–52, 1998
Dog TL: Black cohosh. 2000 In: 7th Annual Course Botanical Medicine in Modern Clinical Practice. New York, May 20-24, 2002, p 556
Boik J: Natural Compounds in Cancer Therapy. Oregon Medical Press, Princeton, 2001, p 304
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Einbond, L.S., Shimizu, M., Xiao, D. et al. Growth Inhibitory Activity of Extracts and Purified Components of Black Cohosh on Human Breast Cancer Cells. Breast Cancer Res Treat 83, 221–231 (2004). https://doi.org/10.1023/B:BREA.0000014043.56230.a3
Issue Date:
DOI: https://doi.org/10.1023/B:BREA.0000014043.56230.a3