Actein and a Fraction of Black Cohosh Potentiate Antiproliferative Effects of Chemotherapy Agents on Human Breast Cancer Cells

Abstract

The purpose of this study was to determine whether the triterpene glycosides present in black cohosh enhance the growth inhibitory effects of specific breast cancer chemotherapy agents. Black cohosh roots and rhizomes were extracted with methanol (MeOH)/water (H₂O) and fractionated by solvent-solvent partitioning to yield three fractions: hexane, ethyl acetate (EtOAc) and water. The EtOAc fraction is enriched in triterpene glycosides, including the compound actein. Actein and the EtOAc fraction were then tested, alone and in combination with chemotherapy agents, for growth inhibition of the ER^- Her2 overexpressing breast cancer cell line MDA-MB-453. We found that actein exerted a synergistic effect on growth inhibition when combined with doxorubicin or 5-flourouracil. Synergy was also obtained when the EtOAc fraction was combined with doxorubicin. Actein increased the percent of cells in the G1 phase of the cell cycle and had a similar effect when combined with 5-flourouracil or doxorubucin. Actein enhanced the induction of apoptosis by paclitaxel, 5-flourouracil or doxorubicin. Our results indicate that relatively low concentrations of actein or the EtOAc fraction of black cohosh can cause synergistic inhibition of human breast cancer cell proliferation when combined with different classes of chemotherapy agents.

Abbreviations

CI:Combination Index

DOX:Doxorubicin

DMEM:Dulbecco's Modified Eagle's medium

FBS:Fetal Bovine Serum

5-FU:5-Fluorouracil

TAX:Paclitaxel

Actaea racemosa - actein - doxorubicin - paclitaxel - Ranunculaceae - synergism - triterpene glycoside

References

• 1 Foster S. Black cohosh: Cimicifuga racemosa. A literature review.  HerbalGram. 1999;  45 35-49
• 2 Rockwell S, Liu Y, Higgins S A. Alteration of the effects of cancer therapy agents on breast cancer cells by the herbal medicine black cohosh.  Breast Cancer Res Treat. 2005;  90 233-9
• 3 Newton K M, Reed S D, Grothaus L, Ehrlich K, Guiltinan J, Ludman E. et al . The herbal alternatives for menopause (HALT) study: background and study design.  Maturitas. 2005;  52 134-46
• 4 Bodinet C, Freudenstein J. Influence of marketed herbal menopause preparations on MCF-7 cell proliferation.  Menopause. 2004;  11 281-9
• 5 Low Dog T, Powell K L, Weisman S M. Critical evaluation of the safety of Cimicifuga racemosa in menopause symptom relief.  Menopause. 2003;  10 299-313
• 6 Borrelli F, Izzo A A, Ernst E. Pharmacological effects of Cimicifuga racemosa .  Life Sci. 2003;  73 1215-29
• 7 Stromeier S, Petereit F, Nahrstedt A. Phenolic esters from the rhizomes of Cimicifuga racemosa do not cause proliferation effects in MCF-7 cells.  Planta Med. 2005;  71 495-500
• 8 Dixon-Shanies D, Shaikh N. Growth inhibition of human breast cancer cells by herbs and phytoestrogens.  Oncol Rep. 1999;  6 1383-7
• 9 Nesselhut T, Schellhase C, Deitrich R, Kuhn W. Studies on mammary carcinoma cells regarding the proliferation potential of herbal medication with estrogen-like effects.  Arch Gynecol Obstet. 1993;  254 817-8
• 10 Bodinet C, Freudenstein J. Influence of Cimicifuga racemosa on the proliferation of estrogen receptor-positive human breast cancer cells.  Breast Cancer Res Treat. 2002;  76 1-10
• 11 Watanbe K, Mimaki Y, Sakagami H, Sashida Y. Cycloartane glycosides from the rhizomes of Cimicifuga racemosa and their cytotoxic activities.  Chem Pharm Bull. 2002;  50 121-5
• 12 Sakurai N, Kozuka M, Tokuda H, Nobukuni Y, Takayasu J, Nishino H. et al . Antitumor agents 220. Antitumor-promoting effects of cimigenol and related compounds on Epstein-Barr virus activation and two-stage mouse skin carcinogenesis.  Bioorg Med Chem. 2003;  11 1137-40
• 13 Hostanska K, Nisslein T, Freudenstein J, Reichling J, Saller R. 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;  84 151-60
• 14 Hostanska K, Nisslein T, Freudenstein J, Saller R. Apoptosis of human prostate androgen-dependent and -independent carcinoma cells induced by an isopropanolic extract of black cohosh involves degradation of cytokeratin (CK) 18.  Anticancer Res. 2005;  25 139-47
• 15 Hostanska K, Nisslein T, Freudenstein J, Reichling J, Saller R. Evaluation of cell death caused by triterpene glycosides and phenolic substances from Cimicifuga racemosa extract in human MCF-7 breast cancer cells.  Biol Pharm Bull. 2004;  27 1970-5
• 16 Einbond L S, Shimizu M, Xiao D, Nuntanakorn P, Lim J T, Suzui M. et al . Growth inhibitory activity of extracts and purified components of black cohosh on human breast cancer cells.  Breast Cancer Res Treat. 2004;  83 221-31
• 17 Chou T C, Talalay P. Qualitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme inhibitors.  Adv Enzyme Regul. 1984;  22 27-55
• 18 Soriano A F, Helfrich B, Chan D C, Heasley L E, Bunn  Jr P A, Chou T C. Synergistic effects of new chemopreventive agents and conventional cytotoxic agents against human lung cancer cell lines.  Cancer Res. 1999;  59 6178-84
• 19 Imoto M, Doki Y, Jiang W, Han E K, Weinstein I B. Effects of cyclin D1 overexpression on G1 progression-related events.  Exp Cell Res. 1997;  236 173-80
• 20 Shimizu M, Deguchi A, Lim J T, Moriwaki H, Kopelovich L, Weinstein I B. (-)-Epigallocatechin gallate and polyphenon E inhibit growth and activation of the epidermal growth factor receptor and human epidermal growth factor receptor-2 signaling pathways in human colon cancer cells.  Cancer Res. 2005;  11 2735-46
• 21 Pegram M, Hsu S, Lewis G, Pietras R, Beryt M, Sliwkowski M. et al . Inhibitory effects of combinations of HER-2/neu antibody and chemotherapeutic agents used for treatment of human breast cancers.  Oncogene. 1999;  18 2241-51
• 22 Yakes F M, Chinratanalab W, Ritter C A, King W, Seelig S, Arteaga C L. Herceptin-induced inhibition of phosphatidylinositol-3 kinase and Akt is required for antibody-mediated effects on p27, cyclin D1, and antitumor action.  Cancer Res . 2002;  62 4132-41
• 23 Nuntanakorn P, Jiang B, Einbond L S, Yang H, Kronenberg F, Weinstein I B. et al . Polyphenolic constituents of Actaea racemosa .  J Nat Prod. 2006;  69 314-8
• 24 Liske E. Therapeutic efficacy and safety of Cimicifuga racemosa for gynecologic disorders.  Adv Ther. 1998;  15 45-53
• 25 Johnson B M, van Breemen R B. In vitro formation of quinoid metabolites of the dietary supplement Cimicifuga racemosa (black cohosh).  Chem Res Toxicol. 2003;  16 838-46
• 26 Hemmi H, Kitame F, Ishida N, Kusano G, Kondo Y, Nozoe S. Inhibition of thymidine transport into phytohemagglutinin-stimulated lymphocytes by triterpenoids from Cimicifuga species.  J Pharmacobiodyn. 1979;  2 339-49
• 27 Tsukamoto S, Aburatani M, Ohta T. Isolation of CYP3A4 inhibitors from the black cohosh (Cimicifuga racemosa).  Evid Based Complement Alternat Med. 2005;  2 223-6