Influence of Rheum rhaponticum, Cimicifuga racemosa and Trifolium pratense extracts on breast cancer cell proliferation

Summary

Background: Extracts from the Siberian rhubarb (Rheum rhaponticum) as well as those from red clover (Trifolium pratense) contain phytoestrogens, while black cohosh (Cimicifuga racemosa) may alternatively modify estrogen receptor activity. They have been used for the treatment of menopausal symptoms. Little is known about their effects on estrogen-dependent tumor cells.

Materials and Methods: The influence of a standardized preparation of Rheum rhaponticum (ERr 731®), a well described preparation of Trifolium pratense (Menoflavon® extra) and a commercial preparation of Cimicifuga racemosa (Cefakliman® mono) on proliferation of breast cancer cells was analyzed by the crystal violet assay in estrogen-receptor (ER) dependent (MCF-7) and ER independent (MDA-MB-231) cell-based systems. As positive control for the proliferation of MCF-7 cells we used 17 β-estradiol. The estrogen specificity was demonstrated by applying the ER antagonist ICI182,780 to the cell culture.

Results: Rheum rhaponticum and Cimicifuga racemosa did not induce cell proliferation of MCF-7 or MDA-MB-231 cells. High concentration (100 µg/mL) of Rheum rhaponticum and Trifolium pratense extract were toxic to both cell lines. Trifolium pratense slightly increased proliferation of MCF-7 cells in a dose dependent manner, but this effect was not estrogen specific.

Conclusion: Our results demonstrate that the used extracts have no estrogen specific impact on the proliferation of ER dependent and ER independent breast cancer cells.

Zusammenfassung

Einfluss von Rheum rhaponticum, Cimicifuga racemosa und Trifolium pratense Extrakten auf die Proliferation von Brustkrebszellen

Hintergrund: Extrakte des Sibirischen Rhabarbers (Rheum rhaponticum) wie auch aus Rotklee (Trifolium pratense) enthalten Phytoöstrogene, während die Traubensilberkerze (Cimicifuga racemosa) möglicherweise auf anderem Weg die Östrogenrezeptoraktivität modifiziert. Sie werden zur Behandlung menopausaler Symptome eingesetzt. Über ihre Effekte auf östrogenabhängige Tumorzellen ist bisher wenig bekannt.

Material und Methoden: Der Einfluss eines standardisierten Rheum-rhaponticum-Präparates (ERr 731®), eines gut charakterisierten Extraktes aus Trifolium pratense (Menoflavon® extra) und eines Handelspräparates aus Cimicifuga racemosa (Cefakliman® mono) auf die Proliferation von Brustkrebszellen wurde in Östrogenrezeptor (ER)-abhängigen (MCF-7) und ER-unabhängigen (MDA-MB-231) zellbasierten Systemen mittels Kristallviolett-Methode untersucht. Als Positivkontrolle für die Proliferation von MCF-7-Zellen wurde 17 β-Estradiol eingesetzt. Die Östrogenspezifität wurde durch Zugabe des ER-Antagonisten ICI182,780 zu der Zellkultur bewiesen.

Ergebnisse: Der Rheum-rhaponticum- und der Cimicifuga-racemosa-Extrakt induzierten keine Proliferation von MCF-7 oder MDA-MB-231-Zellen. Hohe Konzentrationen (100 µg/mL) von Rheum-rhaponticum- und Trifolium-pratense-Extrakt waren für beide Zelllinien toxisch. Trifolium pratense erhöhte die Proliferation von MCF-7 Zellen diskret dosisabhängig, aber dieser Effekt war nicht östrogenspezifisch.

Schlussfolgerung: Unsere Ergebnisse zeigen, dass die verwendeten Extrakte keinen östrogenspezifischen Einfluss auf die Proliferation von ER-abhängigen und ER-unabhängigen Brustkrebszellen haben.

Keywords

Rheum rhaponticum - Cimicifuga racemosa - Trifolium pratense - Phytoestrogens - menopausal symptoms - breast cancer

Schlüsselwörter

Rheum rhaponticum - Cimicifuga racemosa - Trifolium pratense - Phytoöstrogene - menopausale Symptome - Brustkrebs

^a Center for Complementary Medicine, Department of Environmental Health Sciences, University Medical Center Freiburg, Germany

^b Department of Pharmacognosy, University of Vienna, Vienna, Austria

• Literatur

• 1 Hasper I, Ventskovskiy BM, Rettenberger R et al. Long-term efficacy and safety of the special extract ERr 731 of Rheum rhaponticum in perimenopausal women with menopausal symptoms. Menopause N Y N 2009; 16: 117-131
• 2 Heger M, Ventskovskiy BM, Borzenko I et al. Efficacy and safety of a special extract of Rheum rhaponticum (ERr 731) in perimenopausal women with climacteric complaints: a 12-week randomized, double-blind, placebo-controlled trial. Menopause N Y N 2006; 13: 744-759
• 3 Kaszkin-Bettag M, Ventskovskiy BM, Solskyy S et al. Confirmation of the efficacy of ERr 731 in perimenopausal women with menopausal symptoms. Altern Ther Health Med 2009; 15: 24-34
• 4 Burton A. The Bukharans: a dynastic, diplomatic, and commercial history, 1550-1702. New York: St. Martin’s Press; 1997
• 5 Von Linné C, Junk W. Caroli Linnæi ... Species plantarum: exhibentes plantas rite cognitas, ad genera relatas, cum differentiis specificis, nominibus trivialibus, synonymis selectis, locis natalibus, secundum systema sexuale digestas. [Internet]. Impensis Laurentii Salvii 1753
• 6 Wober J, Möller F, Richter T et al. Activation of estrogen receptor-beta by a special extract of Rheum rhaponticum (ERr 731), its aglycones and structurally related compounds. J Steroid Biochem Mol Biol 2007; 107: 191-201
• 7 Rupprich N, Hildebrand H, Kindl H. Substrate specificity in vivo and in vitro in the formation of stilbenes. Biosynthesis of rhaponticin. Arch Biochem Biophys 1980; 200: 72-78
• 8 Kolodziejczyk-Czepas J. Trifolium species-derived substances and extracts − biological activity and prospects for medicinal applications. J Ethnopharmacol 2012; 143: 14-23
• 9 Dixon RA. Phytoestrogens. Annu Rev Plant Biol 2004; 55: 225-261
• 10 Michel T, Halabalaki M, Skaltsounis AL. New concepts, experimental approaches, and dereplication strategies for the discovery of novel phytoestrogens from natural sources. Planta Med 2013; 79: 514-532
• 11 Coon JT, Pittler MH, Ernst E. Trifolium pratense isoflavones in the treatment of menopausal hot flushes: a systematic review and meta-analysis. Phytomedicine 2007; 14: 153-159
• 12 Lethaby AE, Brown J, Marjoribanks J et al. Phytoestrogens for vasomotor menopausal symptoms. Cochrane Database Syst Rev 2007; CD001395-
• 13 Michel T, Halabalaki M, Skaltsounis AL. New concepts, experimental approaches, and dereplication strategies for the discovery of novel phytoestrogens from natural sources. Planta Med 2013; 79: 514-532
• 14 Kuiper GG, Lemmen JG, Carlsson B et al. Interaction of estrogenic chemicals and phytoestrogens with estrogen receptor beta. Endocrinology 1998; 139: 4252-4263
• 15 Tong D, Schuster E, Seifert M et al. Expression of estrogen receptor beta isoforms in human breast cancer tissues and cell lines. Breast Cancer Res Treat 2002; 71: 249-255
• 16 Vladusic EA, Hornby AE, Guerra-Vladusic FK et al. Expression and regulation of estrogen receptor beta in human breast tumors and cell lines. Oncol Rep 2000; 7: 157-167
• 17 Lundholt BK, Briand P, Lykkesfeldt AE. Growth inhibition and growth stimulation by estradiol of estrogen receptor transfected human breast epithelial cell lines involve different pathways. Breast Cancer Res Treat 2001; 67: 199-214
• 18 Petersen OW, Høyer PE, van Deurs B. Frequency and distribution of estrogen receptor-positive cells in normal, nonlactating human breast tissue. Cancer Res 1987; 47: 5748-5751
• 19 Molino A, Micciolo R, Turazza M et al. Estrogen receptors in 699 primary breast cancers: a comparison of immunohistochemical and biochemical methods. Breast Cancer Res Treat 1995; 34: 221-228
• 20 Howell A, Robertson JF, Abram P et al. Comparison of fulvestrant versus tamoxifen for the treatment of advanced breast cancer in postmenopausal women previously untreated with endocrine therapy: a multinational, double-blind, randomized trial. J Clin Oncol 2004; 22: 1605-1613
• 21 Osborne CK, Wakeling A, Nicholson RI. Fulvestrant: an oestrogen receptor antagonist with a novel mechanism of action. Br J Cancer 2004; 90 (Suppl. 1): S2-6
• 22 Püssa T, Raudsepp P, Kuzina K, Raal A. Polyphenolic composition of roots and petioles of Rheum rhaponticum L. Phytochem Anal 2009; 20: 98-103
• 23 Klejdus B, Vacek J, Benesová L et al. Rapid-resolution HPLC with spectrometric detection for the determination and identification of isoflavones in soy preparations and plant extracts. Anal Bioanal Chem 2007; 389: 2277-2285
• 24 Tsao R, Papadopoulos Y, Yang R et al. Isoflavone profiles of red clovers and their distribution in different parts harvested at different growing stages. J Agric Food Chem 2006; 54: 5797-5805
• 25 Li W, Sun Y, Liang W et al. Identification of caffeic acid derivatives in Actea racemosa (Cimicifuga racemosa, black cohosh) by liquid chromatography/tandem mass spectrometry. Rapid Commun Mass Spectrom 2003; 17: 978-982
• 26 Cicek SS, Aberham A, Ganzera M, Stuppner H. Quantitative analysis of cycloartane glycosides in black cohosh rhizomes and dietary supplements by RRLC-ELSD and RRLC-qTOF-MS. Anal Bioanal Chem 2011; 400: 2597-2605
• 27 He K, Pauli GF, Zheng B et al. Cimicifuga species identification by high performance liquid chromatography-photodiode array/mass spectrometric/evaporative light scattering detection for quality control of black cohosh products. J Chromatogr A 2006; 1112: 241-254
• 28 Taxvig C, Elleby A, Sonne-Hansen K et al. Effects of nutrition relevant mixtures of phytoestrogens on steroidogenesis, aromatase, estrogen, and androgen activity. Nutr Cancer 2010; 62: 122-131
• 29 Brzezinski A, Debi A. Phytoestrogens: the “natural” selective estrogen receptor modulators?. Eur J Obstet Gynecol Reprod Biol 1999; 85: 47-51
• 30 Matthews J, Gustafsson JA. Estrogen signaling: a subtle balance between ER alpha and ER beta. Mol Interv 2003; 3: 281-292
• 31 Hajirahimkhan A, Dietz BM, Bolton JL. Botanical modulation of menopausal symptoms: mechanisms of action?. Planta Med 2013; 79: 538-553
• 32 Vollmer G, Papke A, Zierau O. Treatment of menopausal symptoms by an extract from the roots of rhapontic rhubarb: the role of estrogen receptors. Chin Med 2010; 5: 7-
• 33 Keiler AM, Papke A, Kretzschmar G et al. Long-term effects of the rhapontic rhubarb extract ERr 731® on estrogen-regulated targets in the uterus and on the bone in ovariectomized rats. J Steroid Biochem Mol Biol 2012; 128: 62-68
• 34 Papke A, Kretzschmar G, Zierau O et al. Effects of the special extract ERr 731 from Rheum rhaponticum on estrogen-regulated targets in the uterotrophy model of ovariectomized rats. J Steroid Biochem Mol Biol 2009; 117: 176-184
• 35 Al-Akoum M, Dodin S, Akoum A. Synergistic cytotoxic effects of tamoxifen and black cohosh on MCF-7 and MDA-MB-231 human breast cancer cells: an in vitro study. Can J Physiol Pharmacol 2007; 85: 1153-1159
• 36 Gaube F, Wolfl S, Pusch L et al. Gene expression profiling reveals effects of Cimicifuga racemosa (L.) NUTT. (black cohosh) on the estrogen receptor positive human breast cancer cell line MCF-7. BMC Pharmacol 2007; 7: 11-
• 37 Hostanska K, Nisslein T, Freudenstein J et al. Cimicifuga racemosa extract inhibits proliferation of estrogen receptor-positive and negative human breast carcinoma cell lines by induction of apoptosis. Breast Cancer Res Treat 2004; 84: 151-160
• 38 Hostanska K, Nisslein T, Freudenstein J et al. Inhibitory effect of an isopropanolic extract of black cohosh on the invasiveness of MDA-mB 231 human breast cancer cells. Vivo Athens Greece 2007; 21: 349-355
• 39 Pilšáková L, Riečanský I, Jagla F. The physiological actions of isoflavone phytoestrogens. Physiol Res Acad Sci Bohemoslov 2010; 59: 651-664
• 40 An J, Tzagarakis-Foster C, Scharschmidt TC et al. Estrogen receptor beta-selective transcriptional activity and recruitment of coregulators by phytoestrogens. J Biol Chem 2001; 276: 17808-17814
• 41 Tham DM, Gardner CD, Haskell WL. Clinical review 97: Potential health benefits of dietary phytoestrogens: a review of the clinical, epidemiological, and mechanistic evidence. J Clin Endocrinol Metab 1998; 83: 2223-2235
• 42 Geller SE, Shulman LP, van Breemen RB et al. Safety and efficacy of black cohosh and red clover for the management of vasomotor symptoms: a randomized controlled trial. Menopause N Y N 2009; 16: 1156-1166
• 43 Lipovac M, Chedraui P, Gruenhut C et al. The effect of red clover isoflavone supplementation over vasomotor and menopausal symptoms in postmenopausal women. Gynecol Endocrinol 2012; 28: 203-207