Induction of apoptosis in MDA-MB-231 human breast carcinoma cells with an ethanol extract of Cyperus rotundus L. by activating caspases

Park,Sang Eun; Shin,Won Tak; Park,Cheol; Hong,Su Hyun; Kim,Gi-Young; Kim,Sung Ok; Ryu,Chung Ho; Hong,Sang Hoon; Choi,Yung Hyun

doi:10.3892/or.2014.3507

Induction of apoptosis in MDA-MB-231 human breast carcinoma cells with an ethanol extract of Cyperus rotundus L. by activating caspases

Authors:
- Sang Eun Park
- Won Tak Shin
- Cheol Park
- Su Hyun Hong
- Gi-Young Kim
- Sung Ok Kim
- Chung Ho Ryu
- Sang Hoon Hong
- Yung Hyun Choi
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Affiliations: Department of Internal Medicine, Dongeui University College of Korean Medicine, Busan 614-052, Republic of Korea, Department of Molecular Biology, College of Natural Sciences, Dongeui University, Busan 614-714, Republic of Korea, Department of Biochemistry, Dongeui University College of Korean Medicine, Busan 614-052, Republic of Korea, Laboratory of Immunobiology, Department of Marine Life Sciences, Jeju National University, Jeju 690-756, Republic of Korea, Team for Scientification of Korean Medical Intervention (BK21 Plus) and Department of Herbal Pharmacology, Daegu Haany University College of Korean Medicine, Daegu 706-828, Republic of Korea, Division of Applied Life Science (BK 21 Program), Institute of Agriculture and Life Science, Gyeongsang National University, Jinju 660‑701, Republic of Korea
Published online on: September 22, 2014 https://doi.org/10.3892/or.2014.3507
Pages: 2461-2470

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Abstract

Cyperus rotundus L. belongs to the Cyperaceae family and is a well documented traditional medicinal herb. Its rhizome has been reported to possess wide spectrum pharmacological activities including anti-inflammatory and antioxidant activity. However, the cellular and molecular mechanisms of the anticancer activity have not been elucidated. In the present study, we investigated the pro-apoptotic effects of C. rotundu rhizomes in a human breast carcinoma MDA-MB-231 cell model. Treatment of MDA-MB-231 cells with an ethanol extract of C. rotundu rhizomes (EECR) and a methanol extract of C. rotundu rhizomes (MECR), but not a water extract of C. rotundu rhizomes, resulted in potent antiproliferative activity. The activity of the EECR was higher than that of the MECR and was associated with the induction of apoptosis. The induction of apoptosis by the EECR was associated with upregulation of death receptor 4 (DR4), DR5 and pro-apoptotic Bax, as well as downregulation of anti-apoptotic survivin and Bcl-2. EECR treatment also downregulated Bid expression and activated caspase-8 and -9, the respective initiator caspases of the extrinsic and intrinsic apoptotic pathways. The increase in mitochondrial membrane depolarization was correlated with activation of effector caspase-3 and cleavage of poly(ADP-ribose) polymerase, a vital substrate of activated caspase-3. Blockage of caspase activation by pretreatment with a pan-caspase inhibitor consistently inhibited apoptosis and abrogated growth inhibition in EECR-treated MDA-MB-231 cells. Although reactive oxygen species (ROS) increased following treatment with the EECR, inhibiting ROS with a ROS scavenger did not attenuate EECR-induced apoptosis. Furthermore, inhibitors of phosphatidylinositol 3-kinase (PI3K)/Akt and mitogen-activated protein kinase (MAPK) signaling pathways failed to reverse EECR-induced apoptosis and growth inhibition. These results suggest that the pro-apoptotic activity of the EECR may be regulated by a caspase-dependent cascade through activation of both intrinsic and extrinsic signaling pathways that is not associated with ROS generation or the PI3K/Akt and MAPK pathways.

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1	Jemal A, Bray F, Center MM, Ferlay J, Ward E and Forman D: Global cancer statistics. CA Cancer J Clin. 61:69–90. 2011. View Article : Google Scholar
2	Forouzanfar MH, Foreman KJ, Delossantos AM, Lozano R, Lopez AD, Murray CJ and Naghavi M: Breast and cervical cancer in 187 countries between 1980 and 2010: a systematic analysis. Lancet. 378:1461–1484. 2011. View Article : Google Scholar : PubMed/NCBI
3	Senkus E, Kyriakides S, Penault-Llorca F, Poortmans P, Thompson A, Zackrisson S and Cardoso F; ESMO Guidelines Working Group. Primary breast cancer: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow-up. Ann Oncol. 24(Suppl 6): vi7–vi23. 2013. View Article : Google Scholar : PubMed/NCBI
4	Spector D, Deroo LA and Sandler DP: Lifestyle behaviors in black and white women with a family history of breast cancer. Prev Med. 52:394–397. 2011. View Article : Google Scholar : PubMed/NCBI
5	Danial NN and Korsmeyer SJ: Cell death: critical control points. Cell. 116:205–219. 2004. View Article : Google Scholar : PubMed/NCBI
6	Jin Z and El-Deiry WS: Overview of cell death signaling pathways. Cancer Biol Ther. 4:139–163. 2005.PubMed/NCBI
7	Caroppi P, Sinibaldi F, Fiorucci L and Santucci R: Apoptosis and human diseases: mitochondrion damage and lethal role of released cytochrome c as proapoptotic protein. Curr Med Chem. 16:4058–4065. 2009. View Article : Google Scholar : PubMed/NCBI
8	Kaufmann T, Strasser A and Jost PJ: Fas death receptor signalling: roles of Bid and XIAP. Cell Death Differ. 19:42–50. 2012. View Article : Google Scholar : PubMed/NCBI
9	Zha J, Weiler S, Oh KJ, Wei MC and Korsmeyer SJ: Posttranslational N-myristoylation of BID as a molecular switch for targeting mitochondria and apoptosis. Science. 290:1761–1765. 2000. View Article : Google Scholar : PubMed/NCBI
10	Chung KM and Yu SW: Interplay between autophagy and programmed cell death in mammalian neural stem cells. BMB Rep. 46:383–390. 2013. View Article : Google Scholar : PubMed/NCBI
11	Mansilla S, Llovera L and Portugal J: Chemotherapeutic targeting of cell death pathways. Anticancer Agents Med Chem. 12:226–238. 2012. View Article : Google Scholar
12	Rufini A and Melino G: Cell death pathology: the war against cancer. Biochem Biophys Res Commun. 414:445–450. 2011. View Article : Google Scholar : PubMed/NCBI
13	Thebtaranonth C, Thebtaranonth Y, Wanauppathamkul S and Yuthavong Y: Antimalarial sesquiterpenes from tubers of Cyperus rotundus: structure of 10,12-peroxycalamenene, a sesquiterpene endoperoxide. Phytochemistry. 40:125–128. 1995.PubMed/NCBI
14	Zhu M, Luk HH, Fung HS and Luk CT: Cytoprotective effects of Cyperus rotundus against ethanol induced gastric ulceration in rats. Phytother Res. 11:392–394. 1997.
15	Uddin SJ, Mondal K, Shilpi JA and Rahman MT: Antidiarrhoeal activity of Cyperus rotundus. Fitoterapia. 77:134–136. 2006. View Article : Google Scholar
16	Sharma R and Gupta R: Cyperus rotundus extract inhibits acetylcholinesterase activity from animal and plants as well as inhibits germination and seedling growth in wheat and tomato. Life Sci. 80:2389–2392. 2007. View Article : Google Scholar
17	Kilani-Jaziri S, Neffati A, Limem I, Boubaker J, Skandrani I, Sghair MB, Bouhlel I, Bhouri W, Mariotte AM, Ghedira K, Dijoux Franca MG and Chekir-Ghedira L: Relationship correlation of antioxidant and antiproliferative capacity of Cyperus rotundus products towards K562 erythroleukemia cells. Chem Biol Interact. 181:85–94. 2009. View Article : Google Scholar : PubMed/NCBI
18	Yazdanparast R and Ardestani A: In vitro antioxidant and free radical scavenging activity of Cyperus rotundus. J Med Food. 10:667–674. 2007. View Article : Google Scholar : PubMed/NCBI
19	Ardestani A and Yazdanparast R: Cyperus rotundus suppresses AGE formation and protein oxidation in a model of fructose-mediated protein glycoxidation. Int J Biol Macromol. 41:572–578. 2007. View Article : Google Scholar
20	Gupta MB, Palit TK, Singh N and Bhargava KP: Pharmacological studies to isolate the active constituents from Cyperus rotundus possessing anti-inflammatory, anti-pyretic and analgesic activities. Indian J Med Res. 59:76–82. 1971.PubMed/NCBI
21	Seo WG, Pae HO, Oh GS, Chai KY, Kwon TO, Yun YG, Kim NY and Chung HT: Inhibitory effects of methanol extract of Cyperus rotundus rhizomes on nitric oxide and superoxide productions by murine macrophage cell line, RAW 264.7 cells. J Ethnopharmacol. 76:59–64. 2001.PubMed/NCBI
22	Kilani S, Ben Ammar R, Bouhlel I, Abdelwahed A, Hayder N, Mahmoud A, Ghedira K and Chekir-Ghedira L: Investigation of extracts from (Tunisian) Cyperus rotundus as antimutagens and radical scavengers. Environ Toxicol Pharmacol. 20:478–484. 2005. View Article : Google Scholar : PubMed/NCBI
23	Tsoyi K, Jang HJ, Lee YS, Kim YM, Kim HJ, Seo HG, Lee JH, Kwak JH, Lee DU and Chang KC: (+)-Nootkatone and (+)-valencene from rhizomes of Cyperus rotundus increase survival rates in septic mice due to heme oxygenase-1 induction. J Ethnopharmacol. 137:1311–1317. 2011.PubMed/NCBI
24	Lemaure B, Touché A, Zbinden I, Moulin J, Courtois D, Macé K and Darimont C: Administration of Cyperus rotundus tubers extract prevents weight gain in obese Zucker rats. Phytother Res. 21:724–730. 2007.
25	Kumar KH and Khanum F: Hydroalcoholic extract of Cyperus rotundus ameliorates H₂O₂-induced human neuronal cell damage via its anti-oxidative and anti-apoptotic machinery. Cell Mol Neurobiol. 33:5–17. 2013.
26	Hemanth Kumar K, Tamatam A, Pal A and Khanum F: Neuroprotective effects of Cyperus rotundus on SIN-1 induced nitric oxide generation and protein nitration: ameliorative effect against apoptosis mediated neuronal cell damage. Neurotoxicology. 34:150–159. 2013.
27	Fu XC, Shan HL, Bai HB and Hu R: Protective effect of Jiangbaiweiyan tablet on ethanol-induced gastric mucosa injury in rats. Zhejiang Da Xue Xue Bao Yi Xue Ban. 40:391–394. 2011.(In Chinese).
28	He CL, Yi PF, Fan QJ, Shen HQ, Jiang XL, Qin QQ, Song Z, Zhang C, Wu SC, Wei XB, Li YL and Fu BD: Xiang-Qi-Tang and its active components exhibit anti-inflammatory and anticoagulant properties by inhibiting MAPK and NF-κB signaling pathways in LPS-treated rat cardiac microvascular endothelial cells. Immunopharmacol Immunotoxicol. 35:215–224. 2013.PubMed/NCBI
29	Lee SJ, Hwang SO, Noh EJ, Kim DU, Nam M, Kim JH, Nam JH and Hoe KL: Transactivation of bad by vorinostat-induced acetylated p53 enhances doxorubicin-induced cytotoxicity in cervical cancer cells. Exp Mol Med. 46:e762014.
30	Lazebnik YA, Kaufmann SH, Desnoyers S, Poirier GG and Earnshaw WC: Cleavage of poly(ADP-ribose) polymerase by a proteinase with properties like ICE. Nature. 371:346–347. 1994. View Article : Google Scholar : PubMed/NCBI
31	Matés JM, Segura JA, Alonso FJ and Márquez J: Intracellular redox status and oxidative stress: implications for cell proliferation, apoptosis, and carcinogenesis. Arch Toxicol. 82:273–299. 2008.PubMed/NCBI
32	Trueba GP, Sánchez GM and Giuliani A: Oxygen free radical and antioxidant defense mechanism in cancer. Front Biosci. 9:2029–2044. 2004. View Article : Google Scholar : PubMed/NCBI
33	Ashkenazi A and Dixit VM: Apoptosis control by death and decoy receptors. Curr Opin Cell Biol. 11:255–260. 1999. View Article : Google Scholar : PubMed/NCBI
34	Peter ME and Krammer PH: The CD95(APO-1/Fas) DISC and beyond. Cell Death Differ. 10:26–35. 2003. View Article : Google Scholar : PubMed/NCBI
35	Deveraux QL and Reed JC: IAP family proteins - suppressors of apoptosis. Genes Dev. 13:239–252. 1999. View Article : Google Scholar : PubMed/NCBI
36	Roy N, Deveraux QL, Takahashi R, Salvesen GS and Reed JC: The c-IAP-1 and c-IAP-2 proteins are direct inhibitors of specific caspases. EMBO J. 16:6914–6925. 1997. View Article : Google Scholar : PubMed/NCBI
37	Schumacker PT: Reactive oxygen species in cancer cells: live by the sword, die by the sword. Cancer Cell. 10:175–176. 2006. View Article : Google Scholar : PubMed/NCBI
38	Yoeli-Lerner M and Toker A: Akt/PKB signaling in cancer: a function in cell motility and invasion. Cell Cycle. 5:603–605. 2006. View Article : Google Scholar : PubMed/NCBI
39	Hu L, Hofmann J, Lu Y, Mills GB and Jaffe RB: Inhibition of phosphatidylinositol 3′-kinase increases efficacy of paclitaxel in in vitro and in vivo ovarian cancer models. Cancer Res. 62:1087–1092. 2002.
40	Kim EK and Choi EJ: Pathological roles of MAPK signaling pathways in human diseases. Biochim Biophys Acta. 1802:396–405. 2010. View Article : Google Scholar : PubMed/NCBI
41	Wagner EF and Nebreda AR: Signal integration by JNK and p38 MAPK pathways in cancer development. Nat Rev Cancer. 9:537–549. 2009. View Article : Google Scholar : PubMed/NCBI