Pulsatilla chinensis is one of the 50 famous fundamental herbs used in traditional Chinese medicine. Saponins are the main components of P. chinensis. Although the anti-proliferative function of saponins has been established in plenty types of cancer, the role of saponins on tumor invasion and metastasis has not been reported, and the mechanisms of how saponins exert the anti-tumor functions are still poorly characterized. Here, we demonstrate that, in breast cancer (BC) cells, raddeanoside R13, a component of saponins extracted from P. chinensis, exhibits strong anti-proliferative and anti-metastasis ability, accompanied by cell cycle arrest, apoptosis, autophagy, and reversion of epithelial-mesenchymal transition (EMT). Raddeanoside R13 (R13) inhibits BC cell proliferation via the activation of G1/S checkpoint transitions, concomitant with a marked decrease of the positive cell cycle regulators, including cyclin D1, cyclin A, and cyclin B1. R13 induces BC cell apoptosis accompanied by the increased levels of cleaved PARP and caspase-3. R13 inhibits BC cell migration and invasion and regulates the expression of the markers of EMT, which plays a critical role in cancer cell migration and invasion. Moreover, R13 suppresses BC tumor growth and metastasis in nude mice. These data highlight the important role of R13 in BC cell proliferation and progression and suggest that R13 may be a useful drug for BC therapy.
Pulsatilla chinensis Anti-tumor Breast cancer Cell proliferation Invasion Metastasis
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The work was supported by the National Key Subject of Drug Innovation (2011ZX11102), Jiangxi province predominant science and technology innovation project (2010DQB01700), National Natural Science Foundation (81330053, 81272913, 81472589, and 31100604), and Beijing Nova Program (Z141102001814055). Beijing University of Chinese Medicine and Beijing Institute of Biotechnology contributed equally to this work.
Compliance with ethical standards
Animal studies were approved by the Institutional Animal Care Committee of Beijing Institute of Biotechnology.
Conflicts of interest
Liu WK, Ho JC, Cheung FW, et al. Apoptotic activity of betulinic acid derivatives on murine melanoma B16 cell line. Eur J Pharmacol. 2004;49(1–3):71–8.CrossRefGoogle Scholar
Cai Y, Tang YM, Liang BW, et al. Experimental studies on antitumor effects of Pulsatilla chinensis (Bunge) Regel in vitro. Chin Tradit Herbal Drugs. 1999;30(1):441–3.Google Scholar
Xu QM, Shu Z, He WJ, et al. Antitumor activity of Pulsatilla chinensis (Bunge) Regel saponins in human liver tumor 7402 cells in vitro and in vivo. Phytomedicine. 2012;19(3–4):293–300.CrossRefPubMedGoogle Scholar
Liu Q, Chen WC, Jiao Y, et al. Pulsatilla saponin A, an active molecule from Pulsatilla chinensis, induces cancer cell death and inhibits tumor growth in mouse xenograft models. J Surgical research. 2014;188(2):387–95.CrossRefGoogle Scholar
Han LT, Fang Y, Li MM, et al. The antitumor effects of triterpenoid saponins from the anemone flaccida and the underlying mechanism. Evid Based Complement Alternat Med. 2013;517931:2013.Google Scholar
Zheng YT, Zhou F, Wu XL, et al. 23-Hydroxybetulinic acid from Pulsatilla chinensis (Bunge) Regel synergizes the antitumor activities of doxorubicin in vitro and in vivo. J Ethnopharmacology. 2010;128(3):615–22.CrossRefPubMedGoogle Scholar
Xu YJ, Zhou T, Du ZY, et al. A novel animal model established using in vivo imaging to monitor the tumor growth and metastasis. Acta Laboratorium Animalis Scientia Sinica. 2008;16(1):19–22.Google Scholar
Han B, Cui H, Kang L, et al. Metformin inhibits thyroid cancer cell growth, migration, and EMT through the mTOR pathway. Tumour Biol. 2015;36(8):6295–304.CrossRefPubMedGoogle Scholar
Chen J, Chen J, Li Z, et al. The apoptotic effect of apigenin on human gastric carcinoma cells through mitochondrial signal pathway. Tumour Biol. 2014;35(8):7719–26.CrossRefPubMedGoogle Scholar
Hong SW, Jung KH, Lee HS, et al. SB365, Pulsatilla saponin D suppresses the proliferation of human colon cancer cells and induces apoptosis by modulating the AKT/mTOR signalling pathway. Food Chem. 2013;136(1):26–33.CrossRefPubMedGoogle Scholar
Hong SW, Jung KH, Lee HS, et al. SB365 inhibits angiogenesis and induces apoptosis of hepatocellular carcinoma through modulation of PI3K/Akt/mTOR signaling pathway. Cancer Sci. 2012;103(11):1929–37.CrossRefPubMedGoogle Scholar
Hong SW, Jung KH, Lee HS, et al. SB365, Pulsatilla saponin D suppresses proliferation and induces apoptosis of pancreatic cancer cells. Oncolgy reports. 2013;30(2):801–8.Google Scholar
Hong SW, Jung KH, Lee HS, et al. SB365, Pulsatilla saponin D, targets c-Met and exerts antiangiogenic and antitumor activities. Carcinogenesis. 2013;34(9):2156–69.CrossRefPubMedGoogle Scholar
Boland K, Flanagan L, Prehn JH. Paracrine control of tissue regeneration and cell proliferation by Caspase-3. Cell Death Disease. 2013;11, e725.CrossRefGoogle Scholar
Boulares AH, Yakovlev AG, Lvanova V, et al. Role of poly(ADP-ribose) polymerase(PARP) cleavage in apoptosis. Caspase 3-resistant PARP mutant increases rates of apoptosis in transfected cells. J Biol Chem. 1999;274(33):22932–40.CrossRefPubMedGoogle Scholar
Shuo Y, Rui L, Dongkyoo P, et al. Disruption of STAT3 by niclosamide reverses radioresistance of human lung cancer. Mol Cancer Ther. 2014;13(3):606–16.CrossRefGoogle Scholar
Barthomeuf C, Debiton E, Mshvildadze V, et al. In vitro activity of hederacolchisid A1 compared with other saponins from Hedera colchica against proliferation of human carcinoma and melanoma cells. Planta Medcine. 2002;68(8):672–5.CrossRefGoogle Scholar
Sun YX, Liu JC, Yu HT, et al. Isolation and evaluation of immunological adjuvant activities of saponins from the roots of Pulsatilla chinensis with less adverse reactions. Int Immunopharmacol. 2010;10(5):584–90.CrossRefPubMedGoogle Scholar
Nho KJ, Chun JM, Lee AY. Anti-metastatic effects of Rheum Palmatum L. extract in human MDA-MB-231 breast cancer cells. Environ Toxicol Pharmacology, 40(1):30–8, 2015.Google Scholar
Nizamutdinova IT, Lee GW, Lee JS, et al. Tanshinone I suppresses growth and invasion of human breast cancer cells, MDA-MB-231, through regulation of adhesion molecules. Carcinogenesis. 2008;29(10):1885–92.CrossRefPubMedGoogle Scholar
Liu Y, Cao W, Zhang B, et al. The natural compound magnolol inhibits invasion and exhibits potential in human breast cancer therapy. Sci Rep. 2013;3:3098.PubMedPubMedCentralGoogle Scholar
Hao W, Zhang X, Zhao W, et al. Cryptotanshinone induces pro-death autophagy through JNK signaling mediated by reactive oxygen species generation in lung cancer cell. Anticancer Agents Med Chem, 2015, [Epub ahead of print].Google Scholar
Kim N, Jeong S, Jing K. Docosahexaenoic acid induces cell death in human non-small cell lung cancer cells by repressing mTOR via AMPK activation and PI3K/Akt inhibition. Biomed Res Int. 2015;239764.Google Scholar
Levine B, Klionsky DJ. Development by self-digestion: molecular mechanisms and biological functions of autophagy. Dev Cell. 2004;6(4):463–77.CrossRefPubMedGoogle Scholar
Wu M, Xu LY, Wang X, et al. Guttiferone K induces autophagy and sensitizes cancer cells to nutrient stress-induced cell death. Phytomedicine. 2015;22(10):902–10.CrossRefPubMedGoogle Scholar