Anticancer and Anti-metastatic Effects of Supercritical Extracts of Hops (Humulus lupulus L.) and Mango ginger (Curcuma amada Roxb.) in Human Glioblastoma
Keywords:
glycolysis, Warburg effect, hops, mango ginger,, cell migrationAbstract
Glioblastoma is one of the most aggressive, lethal and incurable primary brain tumors with a dismal prognosis in humans. Mango ginger (Curcuma amada) and hops (Humulus lupulus) are two botanicals containing phytochemicals with potential anticancer effects. We have investigated the anticancer and antimetastatic properties of supercritical CO2 extract of mango ginger (CA) and ethanol extract of hops (HL) in the U-87MG human glioblastoma cell line. Both CA and HL individually demonstrate strong cytotoxicity against glioblastoma cells. CompuSyn analysis of cytotoxicity data confirms that CA and HL are synergistic for cytotoxicity with combination index (CI) values of <1.0. Additionally, CA and HL individually as well as the combination significantly inhibit MMP-2 and MMP-9 activity, tumor cell migration (transendothelial cell migration assay) and AKT phosphorylation in U-87MG cells. CA and HL inhibit glycolysis in U-87MG cells as indicated by the inhibition of ATP and lactate synthesis with the CA+HL combination demonstrating strong inhibition of glycolysis via the reduction of ATP and lactate synthesis compared to cells treated by each extract alone. CA and HL treatment down regulates the expression of proteins associated with metastasis, MMP-2 and MMP-9 and up regulates the expression of TIMP1. Proteins associated with apoptosis, inflammation and energy metabolism were also modulated by CA and HL treatment of glioblastoma cells. These results suggest that CA and HL can be combined for the therapeutic management of glioblastomas.
References
. Regina AJ, Demeule M, Laplante A, et al. Multidrug resistance in brain tumors: roles of the blood-brain barrier. Cancer Metastasis Rev 2001; 20: 13-25. [2]. Koetter U, Biendl M. Hops (Humulus lupulus): A review of its historic and medicinal uses. HerbalGram. 2010;87: 44-57. [3]. Karabin M, Hudcova T, Jelinek L, et al. Biotransformations and biological activities of hop flavonoids. Biotechnology Advances 2015;33: 1063-1090. [4]. Overk CR, Yao P, Chadwick LR, et al. Comparison of the in vitro estrogenic activities of compounds from hops (Humulus lupulus) and red clover (Trifolium pratene). J Agric Food Chem 2005;53: 646-653. [5]. Karabin M, Hudcova T, Jelinek L, et al. Biologically active compounds from hops and prospects for their use. Comprehensive Reviews in Food Science and Food Safety 2016;15: 542-566. [6]. Gerhauser C, Alt A, Heiss E, et al. Cancer chemopreventive activity of xanthohumol, a natural product derived from hops. Mol Cancer Ther 2002; 1: 959-969. [7]. Gerhauser C. Beer constituents as potential cancer chemopreventive agents. Eur J Cancer 2005;41: 1941-1954. [8]. Busch C, Noor S, Leischner C, et al. Anti-proliferative activity of hop-derived prenylflavonoids against human cancer cell lines. Wien Med Wochenschr 2015;165: 258-261. [9]. Zolnierczyk AK, Maczka WK, Grabarczyk M, et al. Isoxanthohumol-Biologically active hop flavonoid. Fitotherapia 2015;103: 72-82. [10]. Stevens JF, Miranda CL, Buhler DR, et al. Chemistry and biology of hop flavonoids. J Am Soc Brew Chem 1998;56: 136-145. [11]. De Keukeleire D, De Cooman L, Rong H, et al. Functional properties of hop polyphenols 2: chemistry, biology, pharmacology, ecology. New York, Kluwer Academics/Plenum Publishers1999; 739-760. [12]. Taylor AW, Barofsky E, Kennedy JA, et al. Hop (Humulus lupulus L.) proanthocyanadins characterized by mass spectrometry, acid analysis, and gel permeation chromatography. J Agric Food Chem 2003;51: 4101-4110. [13]. Wohlfart R. Humulus. In: R.Hansel, K. Keller, H. Rimpler and G. Schneider (eds.). Hagers Handbuch der Phrmazeutischen Praxis Vol 5, Ed. 5, pp. 447-458. Belin, Heidelberg: Springer 1993. [14]. Miranda CL, Stevens JF, Helmrich A, et al. Antiproliferative and cytotoxic effects of prenylated flavonoids from hops (Humulus lupulus) in human cancer cell lines. Food Chem Toxicol 1999;37: 271-285. [15]. Steven JF, Page JE. Xanthohumol and related prenylflavonoids from hops and beer: to your good health. Phytochemistry 2004;65: 1317-1330. [16]. Possimiers S, Bolca S, Grootaert C, et al. The prenylflavonoid isoxanthohumol from hops (Humulus lupulus L) is activated into the potent phytoestrogen 8-prenylnaringenin in vitro and in the human intestine. J Nutr 2006;136: 1862-1867. [17]. Ramachandran C, Lollett IV, Escalon E, et al. Anticancer potential and mechanism of action of mango ginger (Curcuma amada Roxb.) supercritical CO2 extract in human glioblastoma cells. J Evid based Complementary Altern Med 2014;20:109-119. [18]. Ramachandran C, Quirin KW, Escalon E, et al. Therapeutic effect of supercritical CO2 extracts of Curcuma species with cancer drugs in rhabdomyosarcoma cell lines. Phytother Res 2015;8:1152-1160. [19]. Ramachandran C, Portalatin G, Quirin KW, et al. Inhibition of AKT signaling by supercritical CO2 extract of mango ginger (Curcuma amada Roxb) in human glioblastoma cells. J Complement Integr Med 2016;12: 307-314. [20]. Ramachandran C, Portalatin G, Prado AM, et al. In vivo antitumor effect of supercritical extracts of mango ginger (Curcuma amada Roxb.) in U-87MG human glioblastoma cells. J Evid based Complementary Altern Med 2016; DOI: 10.1177/2156587216659390. [21]. Chou TC, Talalay P. Analysis of combined drug effects: a new look at an old problem. Trends Pharmacol Sci 1983;4: 450-453. [22]. Kapadia GJ, Rao GS, Ramachandran C, et al. Synergistic cytotoxicity of red beetroot (Beta vulgaris L.) extracts with doxorubicin in human pancreatic, breast and prostate cancer cell lines. J Complement Integr Med 2013;10: 1-10. [23]. Toth M, Fridman R. Assessment of gelatinases (MMP-2 and MMP-9) by gelatin zymography. Methods Mol Med 2012;878: 121-135. [24]. Ramachandran C, Juan A, Quirin KW, et al. Synergistic effect of supercritical CO2 extract of mango ginger with glycolytic inhibitors in human U-87 glioblastoma cell line. J Complement Integr Med 2018; (in press). [25]. Singh S, Hibbaka JJ, Dharmedra S, et al. A bioactive labdane diterpenoid from Curcuma amada and its semisynthetic analogues as antitubercular agents. Eur J Chem 2010;45: 4379-4382. [26]. Sheeja ADB, Nair MS. Facile isolation of (E)-labda-8(17)12-diene-15,16-dial from Curcuma amada and its conversion to other biologically active compounds. Indian J Chem 2014;53: 319-324. [27]. Stevens JF, Ivancic M, Hsu VL, et al. Prenylflavonoids from Humulus lupulus. Phytochemistry 2000;53: 759-75. [28]. Hudcova T, Bryndova J, Fialova K, et al. Antiproliferative effects of prenylflavonoids from hops on human colon cancer cell lines. J Inst Brew 2014;120: 225-230. [29]. Dorn C, Kraus B, Motyl M, et al. Xanthohumol, a chalcon derived from hops, inhibits hepatic inflammation and fibrosis. Mol Nutr Food Res 2010;54: S205-S213. [30]. Desai A, Darland G, Bland JS, et al. META060 attenuates TNF-attenuates TNF--activated inflammation, endothelial-monocyte interactions, and matrix metalloporteinase-9 expression, inhibits NF-B and AP-1inTHP-1 monocytes. J Atherosclerosis 2012;223: 130-136. [31]. Philips N, Samuel M, Arena R, et al. Direct inhibition of elastase and matrix metalloproteinases and stimulation of biosynthesis of fibrillar collagens, elastin, and fibrillins by xanthohumol. J Cosmet Sci 2010;61: 125-132. [32]. Nowak R, Olech M, Nowacka N. Plant polyphenols as chemopreventive agents. In: Watson R, Preedy V, ZibadiS, editors. Polyphenols in human health and disease. San Diego: Elsevier Inc. 2014;1289-1307. [33]. Tan WF, Lin LP, Li MH, et al. Quercetin, a dietary-derived flavonoid, possesses antiangiogenic potential. Eur J Pharmacol 2003;459: 255-262. [34]. Vijayababu MR, Kanagaraj P, Arunkumar A, et al. Quecetin induces p53-dependent apoptosis in human prostate cancer cells by modulating Bcl-2-related proteins: a possible mediation by IGFBP-3. Oncol Res 2006;16: 67-74. [35]. Guruvayoorappan C, Kuttan G. Biophytum sensitivum (L.) DC inhibits tumor cell invasion and metastasis through a mechanism involving regulation of MMPs, prolyl hydroxylase, lysyl oxidase, nm23, ERK-1, ERK-2, STAT-1, and proinflammatory cytokine gene expression in metastatic lung tissue. Integr Cancer Ther 2008;7: 42-50. [36]. Nagase H. Activation mechanisms of matrix metalloproteinases. Biol Chem 1997;378: 151-160. [37]. Birkedal-Hansen H, Moore WG, Bodden MK, Windsor LJ, et al. Matrix metalloproteinases: A review. Crit Rev Oral Biol Med 4. 1993;4: 197-250. [38]. Bellacosa A, Kumar CC, Di Cristfano A, et al. AKT kinases in cancer: implications for therapeutic targeting. Adv Cancer Res 2005;93: 29-86. [39]. Zhao Z, Han F, Yan S, et al. Oxamate-mediated inhibition of lactate dehydrogenase induces protective autophagy in gastric cancer cells: Involvement of the Akt-mTOR pathway. Cancer Lett 2015;358: 17-56. [40]. Zhou M, Zhao Y, Ding Y, et al. Warburg effect in chemosenitivity: targeting lactate dehydrogenase-A re-sensitizes taxol-resitant cancer cells to taxol. Mol Cancer 2010;9: 33.
