Abstract
Fenugreek (Trigonella foenum-graecum) seeds and roots of wild yam (Dioscorea villosa) possess nutritional and medicinal properties and have been used for centuries in traditional medicine to treat different diseases and inflammatory responses. Diosgenin is a natural steroidal sapogenin extracted from fenugreek and wild yam and it is one of the major bioactive compounds used in the treatment of diabetes, hypercholesterolemia, and inflammation. Recent studies have shown a promising effect of diosgenin as an anti-tumor agent for inhibition of cell proliferation and induction of apoptosis in many cancers such as colon cancer, leukemia, breast cancer, and liver cancer. We examined the effects of different concentrations (5, 10, 15, 20, and 25 µM) of diosgenin on proliferation of rat C6 and human T98G glioblastoma cell lines. We noticed that diosgenin had a high inhibitory effect on the growth of both C6 and T98G cell lines. Diosgenin induced the differentiation of glioblastoma cells, as determined by the increase in the expression of the differentiation marker glial fibrillary acidic protein (GFAP); and decreased the dedifferentiation of the cells, as shown by the decrease in the abundance of the dedifferentiation marker proteins Id2, N-Myc, telomerase reverse transcriptase (TERT), and Notch-1. It also induced apoptosis in C6 and T98G cell lines and the molecular mechanisms involved in the induction of apoptosis included increase in pro-apoptotic Bax protein and decrease in anti-apoptotic Bcl-2 protein. Further, the diosgenin-induced suppression of cell migration was correlated with the decrease in expression of matrix metalloproteinase 2 (MMP2) and MMP9; and the inhibition of angiogenesis, as determined by the tube formation assay, was correlated with a decrease in the protein levels of vascular endothelial growth factor (VEGF) and fibroblast growth factor 2 (FGF2). In conclusion, diosgenin showed anti-tumor effects in glioblastoma cells by induction of differentiation and apoptosis and inhibition of migration, invasion, and angiogenesis.
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References
Mentlein R, Forstreuter F, Mehdorn HM, Held-Feindt J (2004) Functional significance of vascular endothelial growth factor receptor expression on human glioma cells. J Neurooncol 67(1-2):9–18. https://doi.org/10.1023/b:neon.0000021737.89357.cc
Brown LB, Eckley M, Wargo KA (2010) A review of glioblastoma multiforme. Oncology 35:3–10
Adamson C, Kanu OO, Mehta AI, Di C, Lin N, Mattox AK, Bigner DD (2009) Glioblastoma multiforme: a review of where we have been and where we are going. Expert Opin Investig Drugs 18:1061–1083. https://doi.org/10.1517/13543780903052764
Ozdemir-Kaynak E, Qutub AA, Yesil-Celiktas O (2018) Advances in glioblastoma multiforme treatment: new models for nanoparticle therapy. Front Physiol 19:170. https://doi.org/10.3389/fphys.2018.00170
Ostrom QT, Gittleman H, Xu J, Kromer C, Wolinsky Y, Kruchko C, Barnholtz-Sloan JS (2016) CBTRUS Statistical report: primary brain and other central nervous system tumors diagnosed in the United States in 2009-2013. Neuro-Oncology 18(5):v1–v75. https://doi.org/10.1093/neuonc/noz150
Shabbeer S, Sobolewski M, Anchoori RK, Kachhap S, Hidalgo M, Jimeno A, Davidson N, Carducci MA, Khan SR (2009) Fenugreek: a naturally occurring edible spice as an anticancer agent. Cancer Biol Ther 8:272–280. https://doi.org/10.4161/cbt.8.3.7443
Sethi G, Shanmugam MK, Warrier S, Merarchi M, Arfuso F, Kumar AP, Bishayee A (2018) Pro-apoptotic and anti-cancer properties of diosgenin: a comprehensive and critical review. Nutrients 10:5. https://doi.org/10.3390/nu10050645
Wani SA, Kumar P (2016) Fenugreek: a review on its nutraceutical properties and utilization in various food products. J Saudi Soc Agric Sci 17:97–106. https://doi.org/10.1016/j.jssas.2016.01007
Raju J, Mehta R (2009) Cancer chemopreventive and therapeutic effects of diosgenin, a food saponin. Nutr Cancer 61:27–35. https://doi.org/10.1080/01635580802357352
Kim JK, Park SU (2018) An update on the biological and pharmacological activities of diosgenin. Excli J. 17:24–28. https://doi.org/10.17179/excli2017-894
Alsemari A, Alkhodairy F, Aldakan A et al (2014) The selective cytotoxic anti-cancer properties and proteomic analysis of Trigonella Foenum-Graecum. BMC Complement Alt Med 14:114. https://doi.org/10.1186/1472-6882-14-114
Son IS, Kim JH, Sohn HY, Son KH, Kim JS, Kwon CS (2007) Antioxidative and hypolipidemic effects of diosgenin, a steroidal saponin of yam (Dioscorea spp.), on high cholesterol fed rats. Biosci Biotechnol Biochem 71:3063–3071. https://doi.org/10.1271/bbb.70472
Ahamad S, Kumar P (2018) Fenugreek: a review on its nutraceutical properties and utilization in various food products. J Saudi Soc Agric Sci 17:97–106
Chen Y, Tang YM, Yu SL, Han YW, Kou JP, Liu BL, Yu BY (2015) Advances in the pharmacological activities and mechanisms of diosgenin. Chin J Nat Med 13(8):578–587. https://doi.org/10.1016/S1875-5364(15)30053-4
Tietze LF, Bell HP, Chandrasekhar S (2003) Natural product hybrids as new leads for drug discovery. Angew Chem Int Ed 42:3996–4028. https://doi.org/10.1002/anie.200200553
Srinivasan S, Koduru S, Kumar R, Venguswamy G, Kyprianou N, Damodaran C (2009) Diosgenin targets Akt-mediated prosurvival signaling in human breast cancer cells. Int J Cancer 125:961–967. https://doi.org/10.1002/ijc.24419
Li F, Fernandez PP, Rajendran P, Hui KM, Sethi G (2010) Diosgenin, a steroidal saponin, inhibits STAT3 signaling pathway leading to suppression of proliferation and chemosensitization of human hepatocellular carcinoma cells. Cancer Lett 292:197–207. https://doi.org/10.1016/j.canlet.2009.12.003
Moalic S, Liagre B, Corbière C, Bianchi A, Dauça M, Bordji K, Beneytout JL (2001) A plant steroid, diosgenin, induces apoptosis, cell cycle arrest and COX activity in osteosarcoma cells. FEBS Lett 506:225–230. https://doi.org/10.1016/s0014-5793(01)02924-6
Das S, Dey KK, Dey G, Pal I, Majumder A, MaitiChoudhury S, Kundu SC, Mandal M (2012) Antineoplastic and apoptotic potential of traditional medicines thymoquinone and diosgenin in squamous cell carcinoma. PLoS ONE 7:e46641. https://doi.org/10.1371/journal.pone.0046641
Chen PS, Shih YW, Huang HC, Cheng HW (2011) Diosgenin, a steroidal saponin, inhibits migration and invasion of human prostate cancer PC-3 cells by reducing matrix metallo proteinase expression. PLoS ONE 6:e20164. https://doi.org/10.1371/journal.pone.0020164
He Z, Chen H, Li G, Zhu H, Gao Y, Zhang L, Sun J (2014) Diosgenin inhibits the migration of human breast cancer MDA-MB-231 cells by suppressing VAV2 activity. Phytomedicine 21:871–876. https://doi.org/10.1016/j.phymed.2014.02.002
Fresno Vara JA, Casado E, de Castro J, Cejas P, Belda-Iniesta C, González-Barón M (2004) PI3K/Akt signaling pathway and cancer. Cancer Treat Rev 30:193–204. https://doi.org/10.1016/j.ctrv.2003.07.007
Shishodia S, Aggarwal BB (2006) Diosgenin inhibits osteoclastogenesis, invasion, and proliferation through the downregulation of Akt, IκB kinase activation and NF-κB-regulated gene expression. Oncogene 25:1463–1473. https://doi.org/10.1038/sj.onc.1209194
Liao WL, Lin JY, Shieh JC, Yeh H, Hsieh Y, Cheng Y, Lee H, Shen C, Cheng C (2019) Induction of G2/M phase arrest by diosgenin via activation of Chk1 kinase and Cdc25C regulatory pathways to promote apoptosis in human breast cancer cells. Int J Mol Sci 21(1):172. https://doi.org/10.3390/ijms21010172
Raju J, Rao CV (2012) Diosgenin, a steroid saponin constituent of yams and fenugreek: emerging evidence for applications in medicine. In: Rasooli I (ed) Bioactive compounds in phytomedicine. InTech, Rijeka, pp 125–142. https://doi.org/10.5772/26700
Salakou S, Kardamakis D, Tsamandas AC, Zolota V, Apostolakis E, Tzelepi V, Papathanasopoulos P, Bonikos DS, Papapetropoulos T, Petsas T, Dougenis D (2007) Increased Bax/Bcl-2 ratio up-regulates caspase-3 and increases apoptosis in the thymus of patients with myasthenia gravis. Vivo 21:123–132
Kim DS, Jeon BK, Lee YE, Woo WH, Mun YJ (2012) Diosgenin Induces Apoptosis in HepG2 cells through generation of reactive oxygen species and mitochondrial pathway. Evidence-based complementary and alternative medicine. 2012:981675. https://doi.org/10.1155/2012/981675
Kroemer G, Petit P, Zamzami N, Vayssière JL, Mignotte B (1995) The biochemistry of programmed cell death. FASEB J 9:1277–1287. https://doi.org/10.1096/fasebj.9.13.7557017
Samejima K, Ogawa H, Ageichik AV, Peterson KL, Kaufmann SH, Kanemaki MT, Earnshaw WC (2014) Auxin-induced rapid degradation of inhibitor of caspase-activated DNase (ICAD) induces apoptotic DNA fragmentation, caspase activation, and cell death: a cell suicide module. J Biol Chem 289:31617–31623. https://doi.org/10.1074/jbc.m114.583542
Liu M, Wang Z, Ju Y, Wong RN, Wu QY (2005) Diosgenin induces cell cycle arrest and apoptosis in human leukemia K562 cells with the disruption of Ca2+ homeostasis. Cancer Chemother Pharmacol 55(1):79–90. https://doi.org/10.1007/s00280-004-0849-3
Lepage C, Léger DY, Bertrand J, Martin F, Beneytout JL, Liagre B (2011) Diosgenin induces death receptor-5 through activation of p38 pathway and promotes TRAIL-induced apoptosis in colon cancer cells. Cancer Lett 301(2):193–202. https://doi.org/10.1016/j.canlet.2010.12.003
Lv L, Zheng L, Dong D, Xu L, Yin L, Xu Y, Qi Y, Han X, Peng J (2013) Dioscin, a natural steroid saponin, induces apoptosis and DNA damage through reactive oxygen species: a potential new drug for treatment of glioblastoma multiforme. Food Chem Toxicol 59:657–669. https://doi.org/10.1016/j.fct.2013.07.012
Xiao L, Guo D, Hu C, Shen W, Shan L, Li C, Liu X, Yang W, Zhang W, He C (2012) Diosgenin promotes oligodendrocyte progenitor cell differentiation through estrogen receptor- mediated ERK1/2 activation to accelerate remyelination. Glia 60:1037–1052. https://doi.org/10.1002/glia.22333
Janardhanan R, Butler JT, Banik NL, Ray SK (2009) N-(4-Hydroxyphenyl) retinamide potentiated paclitaxel for cell cycle arrest and apoptosis in glioblastoma C6 and RG2 cells. Brain Res 1268:142–153. https://doi.org/10.1016/j.brainres.2009.02.064
Wu KJ, Grandori C, Amacker M et al (1999) Direct activation of TERT transcription by c-MYC. Nat Genet 21(2):220–224. https://doi.org/10.1038/6010
Nappez C, Liagre B, Beneytout JL (1995) Changes in lipoxygenase activities in human erythroleukemia (HEL) cells during diosgenin-induced differentiation. Cancer Lett 96:133–140
Mao ZJ, Tang QJ, Zhang CA, Qin ZF, Pang B, Wei PK, Liu B, Chou YN (2012) Anti-proliferation and anti-invasion effects of diosgenin on gastric cancer BGC-823 cells with HIF-1α shRNAs. Int J Mol Sci 13:6521–6533. https://doi.org/10.3390/ijms1305652
Park CM, Park MJ, Kwak HJ, Lee HC, Kim MS, Lee SH, Park IC, Rhee CH, Hong SI (2006) Ionizing radiation enhances matrix metalloproteinase-2 secretion and invasion of glioma cells through Src/epidermal growth factor receptor-mediated p38/Akt and phosphatidylinositol 3-kinase/Akt signaling pathways. Cancer Res 66:8511–8519. https://doi.org/10.1158/0008-5472.can-05-4340
Liu Y, Zheng J, Zhang Y, Wang Z, Yang Y, Bai M, Dai Y (2016) Fucoxanthin activates apoptosis via inhibition of PI3K/Akt/mTOR pathway and suppresses invasion and migration by restriction of p38-MMP-2/9 pathway in human glioblastoma cells. Neurochem Res 41(10):2728–2751. https://doi.org/10.1007/s11064-016-1989-7
Kubiatowski T, Jang T, Lachyankar MB, Salmonsen R, Nabi RR, Quesenberry PJ, Litofsky NS, Ross AH, Recht LD (2001) Association of increased phosphatidylinositol 3-kinase signaling with increased invasiveness and gelatinase activity in malignant gliomas. J Neurosurg 95:480–488. https://doi.org/10.3171/jns.2001
Yu H, Liu Y, Niu C, Cheng Y (2018) Diosgenin increased DDX3 expression in hepatocellular carcinoma. Am J Transl Res 10:3590–3599
Jang HS, Lal S, Greenwood JA (2010) Calpain 2 is required for glioblastoma cell invasion: regulation of matrix metalloproteinase 2. Neurochem Res 35(11):1796–1804. https://doi.org/10.1007/s11064-010-0246-8
Cortesio CL, Chan KT, Perrin BJ, Burton NO, Zhang S, Zhang ZY, Huttenlocher A (2008) Calpain 2 and PTP1B function in a novel pathway with Src to regulate invadopodia dynamics and breast cancer cell invasion. J Cell Biol 180:957–971. https://doi.org/10.1083/jcb.200708048
Kaur B, Khwaja FW, Severson EA, Matheny SL, Brat DJ, Van Meir EG (2005) Hypoxia and the hypoxia-inducible-factor pathway in glioma growth and angiogenesis. Neuro Oncology 7:134–153. https://doi.org/10.1215/s1152851704001115
Wang L, Xue Y, Shen Y, Li W, Cheng Y, Yan X, Shi W, Wang J, Gong Z, Yang G, Guo Chuanliang, Zhou Yiye, Wang Xiang, Zhou Qi, Zeng F (2012) Claudin 6: a novel surface marker for characterizing mouse pluripotent stem cells. Cell Res 22:1082–1085. https://doi.org/10.1038/cr.2012.77
Wang L, Zhang L, Shen W, Liu Y, Luo Y (2016) High expression of VEGF and PI3K in glioma stem cells provides new criteria for the grading of gliomas. Exp Ther Med 11:571–576. https://doi.org/10.3892/etm.2015.2906
Rajesh Y, Biswas A, Das S, Manda M (2017) Diosgenin and temozolamide: a potential combinatorial chemotherapy to overcome temozolamide resistance in glioblastoma multiforme [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4105. https://doi.org/10.1158/1538-7445.am2017-4105
Ignarro RS, Facchini G, de Melo DR, Pelizzaro-Rocha KJ, Ferreira CV, Castilho RF, Rogerio F (2016) Characteristics of sulfasalazine-induced cytotoxicity in C6 rat glioma cells. Neurosci Lett 638:189–195. https://doi.org/10.1016/j.neulet.2016.12.035
Kodera T, Nakagawa T, Kubota T, Kabuto M, Sato K, Kobayashi H (2000) The expression and activation of matrix metalloproteinase-2 in rat brain after implantation of C6 rat glioma cells. J Neurooncol 46(2):105–114
Giakoumettis D, Kritis A, Foroglou N (2018) C6 cell line: the gold standard in glioma research. Hippokratia 3:105–112
Plate KH, Breier G, Millauer B, Ullrich A, Risau W (1993) Up-regulation of vascular endothelial growth factor and its cognate receptors in a rat glioma model of tumor angiogenesis. Cancer Res 53(23):5822–5827
Giard DJ, Aaronson SA, Todaro GJ et al (1973) In vitro cultivation of human tumors: establishment of cell lines derived from a series of solid tumors. J Natl Cancer Inst 51(5):1417–1423. https://doi.org/10.1093/jnci/51.5.1417
Weinmann L, Wischhusen J, Demma MJ, Naumann U, Roth P, Dasmahapatra B, Weller M (2008) A novel p53 rescue compound induces p53-dependent growth arrest and sensitises glioma cells to Apo2L/TRAIL-induced apoptosis. Cell Death Differ 15(4):718–729. https://doi.org/10.1038/sj.cdd.4402301
Dziembowska Magdalena, Danilkiewicz Malgorzata, Wesolowska Aleksandra, Zupanska Agata, Chouaib Salem, Kaminska Bozena (2007) Cross-talk between Smad and p38 MAPK signalling in transforming growth factor β signal transduction in human glioblastoma cells. Biochem Biophys Res Commun 354(4):1101–1106. https://doi.org/10.1016/j.bbrc.2007.01.113
Kiseleva LN, Kartashev AV, Vartanyan NL, Pinevich AA, Samoilovich MP (2016) Characteristics of A172 and T98G cell lines. Tsitologiia 58(5):349–355
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The work was supported in part by the R01 grants (CA-091460 and NS-057811) from the National Institutes of Health (Bethesda, MD, USA).
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Khathayer, F., Ray, S.K. Diosgenin as a Novel Alternative Therapy for Inhibition of Growth, Invasion, and Angiogenesis Abilities of Different Glioblastoma Cell Lines. Neurochem Res 45, 2336–2351 (2020). https://doi.org/10.1007/s11064-020-03093-0
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DOI: https://doi.org/10.1007/s11064-020-03093-0