Preferences help
enabled [disable] Abstract
Number of results
2016 | 63 | 1 | 79-87
Article title

Antiproliferative effect of β-escin - an in vitro study

Title variants
Languages of publication
This study examined the antiproliferative effects of β-escin (E) in cancer cells. The study showed that E inhibited cancer cells growth in a dose-dependent manner. The flow cytometric analysis revealed an escin-induced increase in the sub-G1 DNA content, which is considered to be a marker of apoptosis. Apoptosis was also confirmed by annexin V staining and DNA fragmentation assay. These effects were associated with increased generation of reactive oxygen species (ROS), caspase-3 activation and decreased mitochondrial membrane potential (MMP). Moreover, escin decreased mitochondrial protein content and mitochondrial fluorescence intensity as well as caused depletion of glutathione (GSH). However, activity of glutathione peroxidase (GPx) and glutathione reductase (GR) was not significantly changed in escin-treated cells. In conclusion, our results demonstrated that E has apoptotic effects in human cancer cells through the mechanisms involving mitochondrial perturbation. Although the exact mechanism needs to be investigated further, it can be concluded that E may be a useful candidate agent for cancer treatment.
Physical description
  • Department of Experimental Medicine, Faculty of Medicine, P. J. Šafárik University, 040 11 Košice, Slovak Republic
  • Department of Pharmacology, Faculty of Medicine, P. J. Šafárik University, 040 11 Košice, Slovak Republic
  • Department of Pharmacology, Faculty of Medicine, P. J. Šafárik University, 040 11 Košice, Slovak Republic
  • Department of Medical and Clinical Biochemistry, Faculty of Medicine, P. J. Šafárik University, 040 11 Košice, Slovak Republic
  • Department of Medical and Clinical Biochemistry, Faculty of Medicine, P. J. Šafárik University, 040 11 Košice, Slovak Republic
  • Department of Medical and Clinical Biochemistry, Faculty of Medicine, P. J. Šafárik University, 040 11 Košice, Slovak Republic
  • Department of Pharmacology, Faculty of Medicine, P. J. Šafárik University, 040 11 Košice, Slovak Republic
  • Department of Pharmacology, Faculty of Medicine, P. J. Šafárik University, 040 11 Košice, Slovak Republic
  • Department of Pharmacology, Faculty of Medicine, P. J. Šafárik University, 040 11 Košice, Slovak Republic
  • Armstrong JS, Steinauer KK, Hornung B, Irish JM, Lecane P, Birrell GW, Peehl DM, Knox SJ (2002) Role of glutathione depletion and reactive oxygen species generation in apoptotic signaling in a human B lymphoma cell line. Cell Death Differ 9: 252-263.
  • Bak Y, Ham S, Baatartsogt O, Jung SH, Choi KD, Han TY, Han IY, Yoon DY (2013) A1E inhibits proliferation and induces apoptosis in NCI-H460 lung cancer cells via extrinsic and intrinsic pathways. Mol Biol Rep 40: 4507-4519.
  • Bombardelli E, Morazzoni P (1996) Aesculus hippocastanum L. Fitoterapia 67: 483-511.
  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantitites of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248-254.
  • Ćalić-Dragosavac D, Stevović S, Zdravković-Korać S, Milojević J, Cingel A, Vinterhalter B (2011) Secondary metabolite of horse chestnut in vitro culture. Adv Environ Biol 5: 267-270.
  • Çiftçi GA, Işcan A, Kutlu M (2015) Escin reduces cell proliferation and induces apoptosis on glioma and lung adenocarcinoma cell lines. Cytotechnology 67: 893-904.
  • Circu ML, Aw TY (2010) Reactive oxygen species, cellular redox systems, and apoptosis. Free Radic Biol Med 48: 749-762.
  • Cortassa S, Aon MA, Marbán E, Winslow RL, O'Rourke B (2003) An integrated model of cardiac mitochondrial energy metabolism and calcium dynamics. Biophys J 84: 2734-2755.
  • Devi GS, Prasad MH, Saraswathi I, Raghu D, Rao DN, Reddy PP (2000) Free radicals antioxidant enzymes and lipid peroxidation in different types of leukemias. Clin Chim Acta 293: 53-62.
  • Fleury C, Mignotte B, Vayssiére JL (2002) Mitochondrial reactive oxygen species in cell death signaling. Biochimie 84: 131-141.
  • Floreani M, Petrone M, Debetto P, Palatin P (1997) A comparison between different methods for the determination of reduced and oxidized glutathione in mammalian tissues. Free Radic Res 26: 449-455.
  • Gogvadze V, Orrenius S (2006) Mitochondrial regulation of apoptotic cell death. Chem Biol Interact 163: 4-14.
  • Güney G, Kutlu HM, Işcan A (2013) The apoptotic effects of escin in the H-Ras transformed 5RP7 cell line. Phytother Res 27: 900-905. 900-905.
  • Guo W, Bull Xu B, Yang XW, Liu Q, Cui JR (2003) The anticancer effect of b-escin sodium. Chin Pharmacol 19: 351-352.
  • Hanahan D, Weinberg RA (2011) Hallmarks of cancer: the next generation. Cell 144: 646-674.
  • Harikumar KB, Sung B, Pandey MK, Guha S, Krishnan S, Aggarwal BB (2010) Escin, a pentacyclic triterpene, chemosensitizes human tumor cells through inhibition of nuclear factor-kappaB signaling pathway. Mol Pharmacol 77: 818-827.
  • Heikal AA (2010) Intracellular coenzymes as natural biomarkers for metabolic activities and mitochondrial anomalies. Biomark Med 4: 241-263.
  • Ji DB, Xu B, Liu JT, Ran FX, Cui JR (2011) β-Escin sodium inhibits inducible nitric oxide synthase expression via downregulation of the JAK/STAT pathway in A549 cells. Mol Carcinog 50: 945-960.
  • Kale M, Rathore N, John S, Bhatnagar D (1999) Lipid peroxidative damage on pyrethroid exposure and alterations in antioxidant status in rat erythrocytes: a possible involvement of reactive oxygen species. Toxicol Lett 105: 197-205.
  • Ke N, Wang X, Xu X, Abassi YA (2011) The xCELLigence system for real-time and label-free monitoring of cell viability. Methods Mol Biol 740: 33-43.
  • Khan M, Yi F, Rasul A, Li T, Wang N, Gao H, Gao R, Ma T (2012) Alantolactone induces apoptosis in glioblastoma cells via GSH depletion, ROS generation, and mitochondrial dysfunction. IUBMB Life 64: 783-794.
  • Kirkpatrick ND, Zou C, Brewer MA, Brands WR, Drezek RA, Utzinger U (2005) Endogenous fluorescence spectroscopy of cell suspensions for chemopreventive drug monitoring. Photochem Photobiol 81: 125-134.
  • Küçükkurt I, Ince S, Keleş H, Akkol EK, Avci G, Yeşilada E, Bacak E (2010) Beneficial effects of Aesculus hippocastanum L. seed extract on the body's own antioxidant defense system on subacute administration. J Ethnopharmacol 129: 18-22.
  • Lemar KM, Aon MA, Cortassa S, O'Rourke B, Müller CT, Lloyd D (2007) Diallyl disulphide depletes glutathione in Candida albicans: oxidative stress-mediated cell death studied by two-photon microscopy. Yeast 24: 695-706.
  • Lemasters JJ (1999) V. Necrapoptosis and the mitochondrial permeability transition: shared pathways to necrosis and apoptosis. Am J Physiol 276: G1-G6.
  • Lindner I, Meier C, Url A, Unger H, Grassauer A, Prieschl-Grassauer E, Doerfler P (2010) Beta-escin has potent anti-allergic efficacy and reduces allergic airway inflammation. BMC Immunol 11: 24.
  • Ly JD, Grubb DR, Lawen A (2003) The mitochondrial membrane potential(Δψm) in apoptosis; an update. Apoptosis 8: 115-128.
  • Marí M, Morales A, Colell A, García-Ruiz C, Kaplowitz N, Fernández-Checa JC (2013) Mitochondrial glutathione: features, regulation and role in disease. Biochim Biophys Acta 1830: 3317-3328.
  • Matés JM, Segura JA, Alonso FJ, Márquez J (2012) Oxidative stress in apoptosis and cancer: an update. Arch Toxicol 86: 1649-1665.
  • Mela L, Seitz S (1979) Isolation of mitochondria with emphasis on heart mitochondria from small amounts of tissue. Methods Enzymol 55: 39-46.
  • Mojžišová G, Mojžiš J, Pilátová M, Varinská L, Ivanová L, Strojný L., Richnavský J (2013) Antiproliferative and antiangiogenic properties of horse chestnut extract. Phytother Res 27: 159-165.
  • Mosman T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65: 55-63.
  • Nagata S (2000) Apoptotic DNA fragmentation. Exp Cell Res 256: 12-18.
  • Nemoto S, Takeda K, Yu ZX, Ferrans VJ, Finkel T (2000) Role for mitochondrial oxidants as regulators of cellular metabolism. Mol Cell Biol 20: 7311-7318.
  • Niu YP, Li LD, Wu LM (2008) Beta-aescin: a potent natural inhibitor of proliferation and inducer of apoptosis in human chronic myeloid leukemia K562 cells in vitro. Leuk Lymphoma 49: 1384-1391.
  • Okon IS, Zou MH (2015) Mitochondrial ROS and cancer drug resistance: Implications for therapy. Pharmacol Res 100: 170-174.
  • Olszewska A, Szewczyk A (2013) Mitochondria as a pharmacological target: Magnum overview. IUBMB Life 65: 273-281.
  • Ong PL, Weng BC, Lu FJ, Lin ML, Chang TT, Hung RP, Chen CH (2008) The anticancer effect of protein-extract from Bidens alba in human colorectal carcinoma SW480 cells via the reactive oxidative species- and glutathione depletion-dependent apoptosis. Food Chem Toxicol 46: 1535-1547.
  • Parsons M, Green DR (2010) Mitochondria in cell death. Essays Biochem 47: 99-114.
  • Patlolla JM, Raju J, Swamy MV, Rao CH (2006) β-Escin inhibits colonic aberrant crypt foci formation in rats and regulates the cell cycle growth by inducing p21waf1/cip1 in colon cancer cells. Mol Cancer Ther 5: 1459-1466.
  • Petit PX, Gendron MC, Schrantz N, Métivier D, Kroemer G, Maciorowska Z, Sureau F, Koester S (2001) Oxidation of pyridine nucleotides during Fas- and ceramide-induced apoptosis in Jurkat cells: correlation with changes in mitochondria, glutathione depletion, intracellular acidification and caspase 3 activation. Biochem J 353: 357-367.
  • Pittler MH, Ernst E (2012) Horse chestnut seed extract for chronic venous insufficiency. Cochrane Database Syst Rev 11: CD003230,
  • Pogue BW, Pitts JD, Mycek MA, Sloboda RD, Wilmot CM, Brandsema JF, O'Hara JA (2001) In vivo NADH fluorescence monitoring as an assay for cellular damage in photodynamic therapy. Photochem Photobiol 74: 817-824.
  • Rimmon A, Vexler A, Berkovich L, Earon G, Ron I, Lev-Ari S (2013) Escin chemosensitizes human pancreatic cancer cells and inhibits the nuclear factor-kappab signaling pathway. Biochem Res Int 2013: 251752.
  • Shen DY, Kang JH, Song W, Zhang WQ, Li WG, Zhao Y, Chen QX (2011) Apoptosis of human cholangiocarcinoma cell lines induced by β-escin through mitochondrial caspase-dependent pathway. Phytother Res 25: 1519-1526.
  • Shishodia S, Aggarwal BB (2004) Nuclear factor-kappaB activation mediates cellular transformation, proliferation, invasion angiogenesis and metastasis of cancer. Cancer Treat Res 119: 139-173.
  • Sirtori CR (2001) Aescin: Pharmacology, pharmacokinetics and therapeutic profile. Pharmacol Res 44: 183-193.
  • Suter A, Bommer S, Rechner J (2006) Treatment of patients with venous insufficiency with fresh plant horse chestnut seed extract: a review of 5 clinical studies. Adv Ther 23: 179-190. 10.1007/BF02850359.
  • Tepe B, Tuncer E, Saraydın SU, Ozer H, Sen M, Karadayi K, Inan DS, Karadayi S, Polat Z, Akpulat A, Duman M, Koksal B, Turan M (2013) Antitumoral effects of Allium sivasicum on breast cancer in vitro and in vivo. Mol Biol Rep 40: 597-604.
  • Tomečková V, Štefanišinová M, Bilecová-Rabajdová M, Kriššáková E, Tomečko M, Tóth Š, Pekárová T, Mareková M (2011) A novel method for the detection of liver damage using fluorescence of hepatic mitochondria in a rat model following ischaemia/reperfusion injury of the small intestine. Spectroscopy 26: 237-243.
  • Vidinský B, Gál P, Pilátová M, Vidová Z, Solár P, Varinská L, Ivanová L, Mojžiš J (2012) Anti-proliferative and anti-angiogenic effects of CB2R agonist (JWH-133) in non-small lung cancer cells (A549) and human umbilical vein endothelial cells: an in vitro investigation. Folia Biol (Praha) 58: 75-80.
  • Wang Q, Kuang H, Su Y, Sun Y, Feng J, Guo R, Chan K (2013) Naturally derived anti-inflammatory compounds from Chinese medicinal plants. J Ethnopharmacol 146: 9-39.
  • Wang YW, Wang SJ, Zhou YN, Pan SH, Sun B (2012) Escin augments the efficacy of gemcitabine through down-regulation of nuclear factor-κB and nuclear factor-κB-regulated gene products in pancreatic cancer both in vitro and in vivo. J Cancer Res Clin Oncol 138: 785-797.
  • Wu H, Che X, Zheng Q, Wu A, Pan K, Shao A, Wu Q, Zhang J, Hong Y (2014) Caspases: a molecular switch node in the crosstalk between autophagy and apoptosis. Int J Biol Sci 10: 1072-1083.
  • Zhang X, Yuan L, Yang XW, Jiao SY, Wang Q (2010) Oxidative mechanism for toxicity induced by sodium aescinate in HK-2 cells. Chin J Pharmacol Toxicol 24: 471-475.
  • Zhang Z, Gao J, Cai X, Zhao Y, Wang Y, Lu W, Gu Z, Zhang S, Cao P (2011) Escin sodium induces apoptosis of human acute leukemia Jurkat T cells. Phytother Res 25: 1747-1755.
  • Zhou XY, Fu FH, Li Z, Dong QJ, He J, Wang CH (2009) Escin, a natural mixture of triterpene saponins exhibits antitumor activity against hepatocellular carcinoma. Planta Med 75: 1580-1585.
Document Type
Publication order reference
YADDA identifier
JavaScript is turned off in your web browser. Turn it on to take full advantage of this site, then refresh the page.