Euphol arrests breast cancer cells at the G1 phase through the modulation of cyclin D1, p21 and p27 expression

Wang,Lin; Wang,Guiying; Yang,Dandan; Guo,Xudong; Xu,Yanxin; Feng,Bo; Kang,Jiuhong

doi:10.3892/mmr.2013.1650

Euphol arrests breast cancer cells at the G1 phase through the modulation of cyclin D1, p21 and p27 expression

Authors:
- Lin Wang
- Guiying Wang
- Dandan Yang
- Xudong Guo
- Yanxin Xu
- Bo Feng
- Jiuhong Kang
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Affiliations: Department of Endocrinology, East Hospital, Tongji University School of Medicine, Yangpu, Shanghai 200092, P.R. China, Clinical and Translational Research Center of Shanghai First Maternity and Infant Health Hospital, Tongji University, Yangpu, Shanghai 200092, P.R. China
Published online on: August 22, 2013 https://doi.org/10.3892/mmr.2013.1650
Pages: 1279-1285

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Abstract

Euphorbia tirucalli is a long‑established treatment for a wide variety of cancers. However, the mechanism of its anticancer effect is yet to be elucidated. In the present study, we examined the anticancer effect of euphol, a tetracyclic triterpene alcohol isolated from the sap of Euphorbia tirucalli, in T47D human breast cancer cells. Following the treatment of cells with different doses of euphol for 24, 48 and 72 h, the cell proliferation, cell cycle, and mRNA and protein levels of cell cycle regulatory molecules were analyzed, respectively. Treatment of the cells with euphol resulted in decreased cell viability, which was accompanied by an accumulation of cells in the G1 phase. Further studies demonstrated that euphol treatment downregulated cyclin D1 expression and the hypophosphorylation of Rb. Furthermore, this effect was correlated with the downregulation of cyclin‑dependent kinase 2 (CDK2) expression and the upregulation of the CDK inhibitors p21 and p27. Reduced expression levels of cyclin A and B1 were also observed, corresponding to the decreased distribution of cells in the S and G2/M phases, respectively. These findings indicated that euphol is an active agent in Euphorbia tirucalli that exerts anticancer activity by arresting the cell cycle of cancer cells.

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1	Cancer Genome Atlas Network. Comprehensive molecular portraits of human breast tumours. Nature. 490:61–70. 2012. View Article : Google Scholar : PubMed/NCBI
2	DeSantis C, Siegel R, Bandi P and Jemal A: Breast cancer statistics, 2011. CA Cancer J Clin. 61:409–418. 2011. View Article : Google Scholar
3	de Bono JS, Tolcher AW and Rowinsky EK: The future of cytotoxic therapy: selective cytotoxicity based on biology is the key. Breast Cancer Res. 5:154–159. 2003.PubMed/NCBI
4	Zafonte BT, Hulit J, Amanatullah DF, Albanese C, Wang C, Rosen E, Reutens A, Sparano JA, Lisanti MP and Pestell RG: Cell-cycle dysregulation in breast cancer: breast cancer therapies targeting the cell cycle. Front Biosci. 5:D938–D961. 2000. View Article : Google Scholar : PubMed/NCBI
5	Munagala R, Aqil F and Gupta RC: Promising molecular targeted therapies in breast cancer. Indian J Pharmacol. 43:236–245. 2011. View Article : Google Scholar : PubMed/NCBI
6	Malumbres M and Barbacid M: Cell cycle, CDKs and cancer: a changing paradigm. Nat Rev Cancer. 9:153–166. 2009. View Article : Google Scholar : PubMed/NCBI
7	Schwartz GK and Shah MA: Targeting the cell cycle: a new approach to cancer therapy. J Clin Oncol. 23:9408–9421. 2005. View Article : Google Scholar : PubMed/NCBI
8	Chin YW, Balunas MJ, Chai HB and Kinghorn AD: Drug discovery from natural sources. AAPS J. 8:E239–E253. 2006.PubMed/NCBI
9	Rizk AM, Hammouda FM, El-Missiry MM, Radwan HM and Evans FJ: Biologically active diterpene esters from euphorbia peplus. Phytochemistry. 24:1605–1606. 1985. View Article : Google Scholar
10	Rasool N, Khan AQ and Malik A: A taraxerane type triterpene from Euphorbia tirucalli. Phytochemistry. 28:1193–1195. 1989. View Article : Google Scholar
11	de Melo JG, Santos AG, de Amorim EL, do Nascimento SC and de Albuquerque UP: Medicinal plants used as antitumor agents in Brazil: an ethnobotanical approach. Evid Based Complement Alternat Med. 2011:3653592011.PubMed/NCBI
12	Akihisa T, Ogihara J, Kato J, Yasukawa K, Ukiya M, Yamanouchi S and Oishi K: Inhibitory effects of triterpenoids and sterols on human immunodeficiency virus-1 reverse transcriptase. Lipids. 36:507–512. 2001. View Article : Google Scholar : PubMed/NCBI
13	Akihisa T, Kithsiri Wijeratne EM, Tokuda H, Enjo F, Toriumi M, Kimura Y, Koike K, Nikaido T, Tezuka Y and Nishino H: Eupha-7,9(11),24-trien-3beta-ol (‘antiquol C’) and other triterpenes from Euphorbia antiquorum latex and their inhibitory effects on Epstein-Barr virus activation. J Nat Prod. 65:158–162. 2002.PubMed/NCBI
14	Dutra RC, Claudino RF, Bento AF, Marcon R, Schmidt EC, Bouzon ZL, Pianowski LF and Calixto JB: Preventive and therapeutic euphol treatment attenuates experimental colitis in mice. PLoS One. 6:e271222011. View Article : Google Scholar : PubMed/NCBI
15	Dutra RC, de Souza PR, Bento AF, Marcon R, Bicca MA, Pianowski LF and Calixto JB: Euphol prevents experimental autoimmune encephalomyelitis in mice: evidence for the underlying mechanisms. Biochem Pharmacol. 83:531–542. 2012. View Article : Google Scholar : PubMed/NCBI
16	Passos GF, Medeiros R, Marcon R, Nascimento AF, Calixto JB and Pianowski LF: The role of PKC/ERK1/2 signaling in the anti-inflammatory effect of tetracyclic triterpene euphol on TPA-induced skin inflammation in mice. Eur J Pharmacol. 698:413–420. 2012. View Article : Google Scholar : PubMed/NCBI
17	Yasukawa K, Akihisa T, Yoshida ZY and Takido M: Inhibitory effect of euphol, a triterpene alcohol from the roots of Euphorbia kansui, on tumour promotion by 12-O-tetradecanoylphorbol-13-acetate in two-stage carcinogenesis in mouse skin. J Pharm Pharmacol. 52:119–124. 2000. View Article : Google Scholar
18	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods. 25:402–408. 2001.
19	O’Connor PM, Jackman J, Bae I, Myers TG, Fan S, Mutoh M, Scudiero DA, Monks A, Sausville EA, Weinstein JN, et al: Characterization of the p53 tumor suppressor pathway in cell lines of the National Cancer Institute anticancer drug screen and correlations with the growth-inhibitory potency of 123 anticancer agents. Cancer Res. 57:4285–4300. 1997.
20	Gillett C, Fantl V, Smith R, Fisher C, Bartek J, Dickson C, Barnes D and Peters G: Amplification and overexpression of cyclin D1 in breast cancer detected by immunohistochemical staining. Cancer Res. 54:1812–1817. 1994.PubMed/NCBI
21	Zukerberg LR, Yang WI, Gadd M, Thor AD, Koerner FC, Schmidt EV and Arnold A: Cyclin D1 (PRAD1) protein expression in breast cancer: approximately one-third of infiltrating mammary carcinomas show overexpression of the cyclin D1 oncogene. Mod Pathol. 8:560–567. 1995.
22	Lin SY, Xia W, Wang JC, Kwong KY, Spohn B, Wen Y, Pestell RG and Hung MC: Beta-catenin, a novel prognostic marker for breast cancer: its roles in cyclin D1 expression and cancer progression. Proc Natl Acad Sci USA. 97:4262–4266. 2000. View Article : Google Scholar : PubMed/NCBI
23	Vermeulen K, Van Bockstaele DR and Berneman ZN: The cell cycle: a review of regulation, deregulation and therapeutic targets in cancer. Cell Prolif. 36:131–149. 2003. View Article : Google Scholar : PubMed/NCBI
24	Pagano M, Tam SW, Theodoras AM, Beer-Romero P, Del Sal G, Chau V, Yew PR, Draetta GF and Rolfe M: Role of the ubiquitin-proteasome pathway in regulating abundance of the cyclin-dependent kinase inhibitor p27. Science. 269:682–685. 1995. View Article : Google Scholar : PubMed/NCBI
25	Nigro JM, Baker SJ, Preisinger AC, Jessup JM, Hostetter R, Cleary K, Bigner SH, Davidson N, Baylin S, Devilee P, et al: Mutations in the p53 gene occur in diverse human tumour types. Nature. 342:705–708. 1989. View Article : Google Scholar : PubMed/NCBI
26	Girard F, Strausfeld U, Fernandez A and Lamb NJ: Cyclin A is required for the onset of DNA replication in mammalian fibroblasts. Cell. 67:1169–1179. 1991. View Article : Google Scholar : PubMed/NCBI
27	Resnitzky D, Hengst L and Reed SI: Cyclin A-associated kinase activity is rate limiting for entrance into S phase and is negatively regulated in G1 by p27Kip1. Mol Cell Biol. 15:4347–4352. 1995.PubMed/NCBI
28	Knoblich JA and Lehner CF: Synergistic action of Drosophila cyclins A and B during the G2-M transition. EMBO J. 12:65–74. 1993.PubMed/NCBI
29	Li H, Sun L, Tang Z, Fu L, Xu Y, Li Z, Luo W, Qiu X and Wang E: Overexpression of TRIM24 correlates with tumor progression in non-small cell lung cancer. PLoS One. 7:e376572012. View Article : Google Scholar : PubMed/NCBI
30	Darzynkiewicz Z, Bruno S, Del Bino G, Gorczyca W, Hotz MA, Lassota P and Traganos F: Features of apoptotic cells measured by flow cytometry. Cytometry. 13:795–808. 1992. View Article : Google Scholar : PubMed/NCBI
31	Nieves-Neira W and Pommier Y: Apoptotic response to camptothecin and 7-hydroxystaurosporine (UCN-01) in the 8 human breast cancer cell lines of the NCI Anticancer Drug Screen: multifactorial relationships with topoisomerase I, protein kinase C, Bcl-2, p53, MDM-2 and caspase pathways. Int J Cancer. 82:396–404. 1999. View Article : Google Scholar