Journal of Cancer Research and Treatment
ISSN (Print): 2374-1996 ISSN (Online): 2374-2003 Website: Editor-in-chief: Jean Rommelaere
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Journal of Cancer Research and Treatment. 2016, 4(4), 61-68
DOI: 10.12691/jcrt-4-4-2
Open AccessArticle

Oleuropein is a Powerful Sensitizer of Doxorubicin-mediated Killing of Prostate Cancer Cells and Exerts Its Action via Induction of Autophagy

Anastasia Papachristodoulou1, 2, Magafoula Tsoukala1, 2, Dimitra Benaki2, Sarantos Kostidis2, Katerina Gioti1, Nektarios Aligiannis3, Harris Pratsinis4, Dimitris Kletsas4, Alexios-Leandros Skaltsounis3, Emmanuel Mikros2, and Roxane Tenta1,

1Department of Nutrition Science and Dietetics, Harokopio University, Athens, Greece

2Division of Pharmaceutical Chemistry, Faculty of Pharmacy, National and Kapodistrian University of Athens, Greece

3Division of Pharmacognosy and Chemistry of Natural Products, Faculty of Pharmacy, National and Kapodistrian University of Athens, Greece

4Laboratory of Cell Proliferation and Ageing, Institute of Biosciences & Applications, National Centre for Scientific Research "Demokritos", Athens, Greece

Pub. Date: August 06, 2016

Cite this paper:
Anastasia Papachristodoulou, Magafoula Tsoukala, Dimitra Benaki, Sarantos Kostidis, Katerina Gioti, Nektarios Aligiannis, Harris Pratsinis, Dimitris Kletsas, Alexios-Leandros Skaltsounis, Emmanuel Mikros and Roxane Tenta. Oleuropein is a Powerful Sensitizer of Doxorubicin-mediated Killing of Prostate Cancer Cells and Exerts Its Action via Induction of Autophagy. Journal of Cancer Research and Treatment. 2016; 4(4):61-68. doi: 10.12691/jcrt-4-4-2


The phenolic component Oleuropein (OLEU), a bioactive natural product, has recently shown antiproliferative properties. Doxorubicin (DXR) is an anthracycline present in many chemotherapeutic schemes, although limited due to its cardio-toxic effects. Important research effort has been devoted therefore, to reducing DXR toxicity without compromising its antitumor efficacy. The anticancer actions of DXR and OLEU were assessed, on PC-3 prostate cancer cells, while cell cycle distribution and rate of apoptosis were assessed by flow cytometry. The autophagic process was determined via immunoblotting and immunofluorescent staining. Finally, cell extracts were analyzed by NMR spectroscopy. The present study showed that both DXR and OLEU inhibited PC-3 cells proliferation, while the co-treatment with DXR and OLEU resulted in an additive inhibition. Although the addition of OLEU to DXR did not alter significantly the cell cycle distribution, exhibited by each treatment alone, and produced a marginal increase on the rate of apoptosis, both compounds produced a remarkable induction of autophagy. In addition, treated cells exhibited significant metabolite alterations. This study demonstrates that OLEU, a basic component of the everyday diet, is capable of lowering significantly the cytotoxic dose of DXR, while obtaining an important anti-proliferative effect in prostate cancer cells.

oleuropein doxorubicin autophagy prostate metabolic profiling

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[1]  Keys A, A.C., Van Buchem FSP The diet and all-causes death rate in the Seven Countries Study. Lancet, 1981. 2(8237): p. 58-61.
[2]  Boring, C.C., T.S. Squires, and T. Tong, Cancer statistics, 1992. CA Cancer J Clin, 1992. 42(1): p. 19-38.
[3]  Minotti, G., et al., Anthracyclines: molecular advances and pharmacologic developments in antitumor activity and cardiotoxicity. Pharmacol Rev, 2004. 56(2): p. 185-229.
[4]  Owen, R.W., et al., Olive-oil consumption and health: the possible role of antioxidants. Lancet Oncol, 2000. 1: p. 107-12.
[5]  Tripoli, E., et al., The phenolic compounds of olive oil: structure, biological activity and beneficial effects on human health. Nutr Res Rev, 2005. 18(1): p. 98-112.
[6]  Goldsmith, C.D., et al., Phytochemical properties and anti-proliferative activity of Olea europaea L. leaf extracts against pancreatic cancer cells. Molecules, 2015. 20(7): p. 12992-3004.
[7]  Fayyaz, S., et al., Oleuropein mediated targeting of signaling network in cancer. Curr Top Med Chem, 2016. 16.
[8]  Andreadou, I., et al., The olive constituent oleuropein exhibits anti-ischemic, antioxidative, and hypolipidemic effects in anesthetized rabbits. J Nutr, 2006. 136(8): p. 2213-9.
[9]  Andreadou, I., et al., Acute doxorubicin cardiotoxicity is successfully treated with the phytochemical oleuropein through suppression of oxidative and nitrosative stress. J Mol Cell Cardiol, 2007. 42(3): p. 549-58.
[10]  DiPaola, R.S., et al., Therapeutic starvation and autophagy in prostate cancer: a new paradigm for targeting metabolism in cancer therapy. Prostate, 2008. 68(16): p. 1743-52.
[11]  Mathew, R., V. Karantza-Wadsworth, and E. White, Role of autophagy in cancer. Nat Rev Cancer, 2007. 7(12): p. 961-7.
[12]  Shintani, T. and D.J. Klionsky, Autophagy in health and disease: a double-edged sword. Science, 2004. 306(5698): p. 990-5.
[13]  Kabeya, Y., et al., LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. EMBO J, 2000. 19(21): p. 5720-8.
[14]  Zoidou, E., et al., Identification of Throuba Thassos, a traditional Greek table olive variety, as a nutritional rich source of oleuropein. J Agric Food Chem, 2010. 58(1): p. 46-50.
[15]  Kaighn, M.E., et al., Establishment and characterization of a human prostatic carcinoma cell line (PC-3). Invest Urol, 1979. 17(1): p. 16-23.
[16]  Vistica, D.T., et al., Tetrazolium-based assays for cellular viability: a critical examination of selected parameters affecting formazan production. Cancer Res, 1991. 51(10): p. 2515-20.
[17]  Le Belle, J.E., et al., A comparison of cell and tissue extraction techniques using high-resolution 1H-NMR spectroscopy. NMR Biomed, 2002. 15(1): p. 37-44.
[18]  Pechlivanis, A., et al., (1)H NMR-based metabonomic investigation of the effect of two different exercise sessions on the metabolic fingerprint of human urine. J Proteome Res, 2010. 9(12): p. 6405-16.
[19]  Tenta, R., et al., Bone microenvironment-related growth factors modulate differentially the anticancer actions of zoledronic acid and doxorubicin on PC-3 prostate cancer cells. Prostate, 2004. 59(2): p. 120-31.
[20]  Kapadia, G.J., et al., Synergistic cytotoxicity of red beetroot (Beta vulgaris L.) extract with doxorubicin in human pancreatic, breast and prostate cancer cell lines. J Complement Integr Med, 2013. 10: p. 113-122.
[21]  Das, A., et al., Sildenafil increases chemotherapeutic efficacy of doxorubicin in prostate cancer and ameliorates cardiac dysfunction. Proc Natl Acad Sci U S A, 2010. 107(42): p. 18202-7.
[22]  Katsoulieris, E.N., The olive leaf extract oleuropein exerts protective effects against oxidant-induced cell death, concurrently displaying pro-oxidant activity in human hepatocarcinoma cells. Redox Rep, 2016. 21(2): p. 90-7.
[23]  Giner, E., et al., Chemopreventive effect of oleuropein in colitis-associated colorectal cancer in c57bl/6 mice. Mol Nutr Food Res, 2016. 60(2): p. 242-55.
[24]  Cheng, J.S., et al., The effect of oleuropein from olive leaf (Olea europaea) extract on Ca(2+) homeostasis, cytotoxicity, cell cycle distribution and ROS signaling in HepG2 human hepatoma cells. Food Chem Toxicol, 2016. 91: p. 151-66.
[25]  Acquaviva, R., et al., Antiproliferative effect of oleuropein in prostate cell lines. Int J Oncol, 2012. 41(1): p. 31-8.
[26]  Hippert, M.M., P.S. O'Toole, and A. Thorburn, Autophagy in cancer: good, bad, or both? Cancer Res, 2006. 66(19): p. 9349-51.
[27]  Singletary, K. and J. Milner, Diet, autophagy, and cancer: a review. Cancer Epidemiol Biomarkers Prev, 2008. 17(7): p. 1596-610.
[28]  Kaur, J. and J. Debnath, Autophagy at the crossroads of catabolism and anabolism. Nat Rev Mol Cell Biol, 2015. 16(8): p. 461-72.
[29]  Ortega, A.L., S. Mena, and J.M. Estrela, Glutathione in cancer cell death. Cancers (Basel), 2011. 3(1): p. 1285-310.
[30]  Jorgenson, T.C., W. Zhong, and T.D. Oberley, Redox imbalance and biochemical changes in cancer. Cancer Res, 2013. 73(20): p. 6118-23.
[31]  Kawaguchi, M., et al., Autophagy is an important metabolic pathway to determine leukemia cell survival following suppression of the glycolytic pathway. Biochem Biophys Res Commun, 2016. 474(1): p. 188-92.
[32]  Lilienbaum, A., Relationship between the proteasomal system and autophagy. Int J Biochem Mol Biol, 2013. 4(1): p. 1-26.
[33]  Ding, W.X., et al., Linking of autophagy to ubiquitin-proteasome system is important for the regulation of endoplasmic reticulum stress and cell viability. Am J Pathol, 2007. 171(2): p. 513-24.
[34]  Copeland, R.J., J.W. Bullen, and G.W. Hart, Cross-talk between GlcNAcylation and phosphorylation: roles in insulin resistance and glucose toxicity. Am J Physiol Endocrinol Metab, 2008. 295(1): p. E17-28.
[35]  de Queiroz, R.M., E. Carvalho, and W.B. Dias, O-GlcNAcylation: The Sweet Side of the Cancer. Front Oncol, 2014. 4: p. 132.
[36]  Munkley, J., I.G. Mills, and D.J. Elliott, The role of glycans in the development and progression of prostate cancer. Nat Rev Urol, 2016. 13(6): p. 324-33.
[37]  Itkonen, H.M., et al., O-GlcNAc transferase integrates metabolic pathways to regulate the stability of c-MYC in human prostate cancer cells. Cancer Res, 2013. 73(16): p. 5277-87.
[38]  Leri, M., et al., The polyphenol Oleuropein aglycone hinders the growth of toxic transthyretin amyloid assemblies. J Nutr Biochem, 2016. 30: p. 153-66.
[39]  Potocnjak, I., et al., Oral administration of oleuropein attenuates cisplatin-induced acute renal injury in mice through inhibition of ERK signaling. Mol Nutr Food Res, 2016. 60(3): p. 530-41.