Please wait a minute...
 Home  About the Journal Editorial Board Aims & Scope Peer Review Policy Subscription Contact us
 
Early Edition  //  Current Issue  //  Open Special Issues  //  Archives  //  Most Read  //  Most Downloaded  //  Most Cited
Aging and Disease    2015, Vol. 6 Issue (6) : 456-465     DOI: 10.14336/AD.2015.0327
Original Article |
Genistein, the Isoflavone in Soybean, Causes Amyloid Beta Peptide Accumulation in Human Neuroblastoma Cell Line: Implications in Alzheimer's Disease
Gargi Chatterjee,Debashree Roy,Vineet Kumar Khemka,Mrittika Chattopadhyay,Sasanka Chakrabarti*
Department of Biochemistry, Institute of Post Graduate Medical Education and Research, 244 AJC Bose Road, Kolkata-700020, India.
Download: PDF(1529 KB)   HTML
Export: BibTeX | EndNote | Reference Manager | ProCite | RefWorks     Supporting Info
Guide   
Abstract  

The isoflavone, genistein, present in soybean is being actively investigated for its potential beneficial effect against Alzheimer’s disease. Our data, however, show that in SHSY5Y cells genistein causes increased expression (mRNA and protein) of amyloid precursor protein (APP), increased mRNA expression and activity of β-secretase and diminished level of insulin degrading enzyme (IDE) which also degrades amyloid beta peptide. These effects of genistein lead to enhanced accumulation of amyloid beta peptide (Aβ42) in SHSY5Y cells. The results do not support the view that genistein could be a putative drug against AD and instead strengthen the epidemiological study which implies that genistein content of soybean food product (Tofu) leads to cognitive impairment.
Keywords Alzheimer’s disease      genistein      amyloid precursor protein      β-secretase      insulin degrading enzyme      amyloid β42.     
ZTFLH:     
Fund: 
Just Accepted Date: 03 April 2015   Issue Date: 18 November 2015
Service
E-mail this article
E-mail Alert
RSS
Articles by authors
Gargi Chatterjee
Debashree Roy
Vineet Kumar Khemka
Mrittika Chattopadhyay
Sasanka Chakrabarti
Cite this article:   
Gargi Chatterjee,Debashree Roy,Vineet Kumar Khemka, et al. Genistein, the Isoflavone in Soybean, Causes Amyloid Beta Peptide Accumulation in Human Neuroblastoma Cell Line: Implications in Alzheimer's Disease[J]. A&D, 2015, 6(6): 456-465.
URL:  
http://www.aginganddisease.org/EN/10.14336/AD.2015.0327     OR     http://www.aginganddisease.org/EN/Y2015/V6/I6/456
Figure 1.  Changes in APP expression by genistein in SHSY5Y cells. SHSY5Y cells were incubated without (control) or with varying concentrations of genistein (10 - 30 µM) for 48 h followed by the measurement of APP mRNA expressions or intracellular level of APP as described in the MATERIALS AND METHODS. (A) RT-qPCR analysis of APP mRNA expression. (B) Representative immunoblot of APP in control or 20 µM genistein-treated cells. The two lanes were aligned from different parts of the same blot. C. Densitometric analysis of APP band intensity. The expression level was presented as fold increase over control. The results are the means ± SEM of 6 observations. Statistically significant, Ψp< 0.05 vs. control.
Figure 2.  β-Secretase expression and activity in SHSY5Y cells after exposure to genistein. The SHSY5Y cells were incubated without (control) or with varying concentrations of genistein (10 - 30 µM) for 48 h. The cell lysate was analyzed for β-secretase mRNA expression or activity as described in the text. (A) RT-qPCR analysis of β-secretase mRNA expression. The results, expressed as fold increase over control, are the means ± SEM of 4 observations. Statistically significant, Ψp< 0.05 vs. control. (B) β-Secretase enzyme activity. The results are the means ± SEM of 6 observations. Statistically significant, ϕp< 0.01 vs. control.
Figure 3.  Genistein effect on IDE and neprilysin level in SHSY5Y cells. SHSY5Y cells were grown in the absence (control) or in the presence of genistein (20 µM) for 48 h. The cell lysate was analysed for IDE content by immunoblotting using γ-actin as the loading control or neprilysin enzyme activity as described in the ‘MATERIALS AND METHODS’. (A) Representative blot. (B) Densitometric analysis of IDE immunoblots with the band intensity expressed as the fold changes with respect to the control. C. Neprilysin enzyme activity. The results are the means ± SEM of 6 observations. Statistically significant, *p< 0.001 vs. control; ϕp< 0.01 vs. control.
Figure 4.  Effect of genistein on Aβ42 content of SHSY5Y cells. SHSY5Y cells were incubated without (control) or with 20 µM of genistein for 48 h. (A) The cell lysate was analyzed for Aβ oligomers by immunoblotting as mentioned in the text. (B) Amido black stained membrane from a parallel blot. (C) Oligomerized Aβ42 analyzed by 5 - 15% SDS-PAGE followed by immunoblotting.
Figure 5.  Immunoassay of Aβ42. Amyloid β42 in the cell lysate as well as that released in the culture medium was measured as described in ‘MATERIALS AND METHODS’ from control and genistein (20 µM) treated SHSY5Y cells. A. Released Aβ42 (extracellular) in culture supernatant. B. Intracellular Aβ42 in cell lysate. The results are the mean ± SEM of four experimental observations. Statistically significant, ϕp< 0.01 vs. control; Ψp< 0.05 vs. control.
[1] Fukutake M, Takahashi M, Ishida K, Kawamura H, Sugimura T, Wakabayashi K (2000). Quantification of genistein and genistein in soybeans and soybean products. Food Chem Toxicol, 34: 457-61.
[2] Omoni AO, Aluko RE (2005). Soybean foods and their benefits: potential mechanisms of action. Nutr Rev, 63: 272-83.
http://dx.doi.org/10.1111/j.1753-4887.2005.tb00141.x
[3] Yuan B, Wang L, Jin Y, Zhen H, Xu P, Xu Y, Li C, Xu H (2012). Role of metabolism in the effects of genistein and its phase II conjugates on the growth of human breast cell lines. AAPS J, 14: 329-44.
http://dx.doi.org/10.1208/s12248-012-9338-5
[4] George J, Banik NL, Ray SK (2010). Genistein induces receptor and mitochondrial pathways and increases apoptosis during BCL-2 knockdown in human malignant neuroblastoma SK-N-DZ cells. J Neurosci Res, 88: 877-86.
[5] Correia SC, Santos RX, Santos MS, Casadesus G, Lamanna JC, Perry G, Smith MA, Moreira PI (2013). Mitochondrial abnormalities in a streptozotocin-induced rat model of sporadic Alzheimer's disease. Curr Alzheimer Res, 10: 406-19.
http://118.145.16.217/magsci/article/article?id=19720439
[6] Kreijkamp-Kaspers S, Kok L, Grobbee DE, de Haan EH, Aleman A, Lampe JW, van der Schouw YT (2004). Effect of soy protein containing isoflavones on cognitive function, bone mineral density, and plasma lipids in postmenopausal women: a randomized controlled trial. JAMA, 292: 65-74.
[7] Bagheri M, Joghataei MT, Mohseni S, Roghani M (2011). Genistein ameliorates learning and memory deficits in amyloid β (1-40) rat model of Alzheimer's disease. Neurobiol Learn Mem, 95: 270-76.
http://118.145.16.217/magsci/article/article?id=15239518
[8] Baluchnejadmojarad T (2009). The Effect of Genistein on Intracerebroventricular Streptozotocin-Induced Cognitive Deficits in Male Rat. Basic Clin Neurosci, (Autumn), 1: 17-21.
[9] Valles SL, Dolz-Gaiton P, Gambini J, Borras C, Lloret A, Pallardo FV, Viña J (2010). Estradiol or genistein prevent Alzheimer's disease-associated inflammation correlating with an increase PPAR gamma expression in cultured astrocytes. Brain Res, 1312:138-44.
http://dx.doi.org/10.1016/j.brainres.2009.11.044
[10] Bang OY, Hong HS, Kim DH, Kim H, Boo JH, Huh K, Mook-Jung I (2004). Neuroprotective effect of genistein against beta amyloid-induced neurotoxicity. Neurobiol Dis, 16: 21-28.
http://118.145.16.217/magsci/article/article?id=14684418
[11] Bagheri M, Rezakhani A, Nyström S, Turkina MV, Roghani M,et al. (2013). Amyloid Beta1-40-induced astrogliosis and the effect of genistein treatment in rat: a three-dimensional confocal morphometric and proteomic study. PLoS ONE, 8: e76526.
http://dx.doi.org/10.1371/journal.pone.0076526
[12] Zeng H, Chen Q, Zhao B (2004). Genistein ameliorates beta-amyloid peptide (25-35)-induced hippocampal neuronal apoptosis. Free Radic Biol Med, 36: 180-88.
http://dx.doi.org/10.1016/j.freeradbiomed.2003.10.018
[13] Viña J. Genistein as a possible treatment for Alzheimer's disease. Valencia, Spain. NCT01982578
[14] White LR, Petrovitch H, Ross GW, Masaki K, Hardman J, Nelson J, Davis D, Markesbery W (2000). Brain aging and midlife tofu consumption. J Am Coll Nutr, 19: 242-55.
http://dx.doi.org/10.1080/07315724.2000.10718923
[15] Hogervorst E, Sadjimim T, Yesufu A, Kreager P, Rahardjo TB (2008). High tofu intake is associated with worse memory in elderly Indonesian men and women. Dement Geriatr Cogn Disord, 26: 50-57.
http://dx.doi.org/10.1159/000141484
[16] Kim H, Peterson TG, Barnes S (1998). Mechanisms of action of the soy isoflavone genistein: emerging role for its effects via transforming growth factor beta signaling pathways. Am J Clin Nutr, 68(6 Suppl): 1418S-25S.
[17] Xie X, Wang SS, Wong TC, Fung MC (2013). Genistein promotes cell death of ethanol-stressed HeLa cells through the continuation of apoptosis or secondary necrosis. Cancer Cell Int, 13:63.
http://dx.doi.org/10.1186/1475-2867-13-63
[18] Dang ZC, Audinot V, Papapoulos SE, Boutin JA, Löwik CW (2003). Peroxisome proliferator-activated receptor gamma (PPARgamma) as a molecular target for the soy phytoestrogen genistein. J Biol Chem, 278: 962-67.
http://dx.doi.org/10.1074/jbc.M209483200
[19] Webster NJ, Green KN, Peers C, Vaughan PF (2002). Altered processing of amyloid precursor protein in the human neuroblastoma SH-SY5Y by chronic hypoxia. J Neurochem, 83: 1262-71.
http://dx.doi.org/10.1046/j.1471-4159.2002.01236.x
[20] Huang H, Bihaqi SW, Cui L, Zawia NH (2011). In vitro Pb exposure disturbs the balance between Aβ production and elimination: the role of AβPP and neprilysin. Neurotoxicology, 32: 300-06.
http://118.145.16.217/magsci/article/article?id=15249766
[21] Cheung YT, Lau WK, Yu MS, Lai CS, Yeung SC, So KF, Chang RC (2009). Effects of all-trans-retinoic acid on human SH-SY5Y neuroblastoma as in vitro model in neurotoxicity research. Neurotoxicology, 30: 127-35.
http://118.145.16.217/magsci/article/article?id=15277243
[22] Tundo GR, Sbardella D, Ciaccio C, Bianculli A, Orlandi A, Desimio MG, Arcuri G, Coletta M, Marini S (2013). Insulin-degrading enzyme (IDE): a novel heat shock-like protein. J Biol Chem, 288: 2281-89.
http://118.145.16.217/magsci/article/article?id=20050204
[23] Page M & Thorpe R (2002). Protein blotting by electroblotting. In J.M. Walker(Ed.). The Protein Protocols Handbook(pp. 317-319). New Jersey: Humana Press.
[24] Sinha M, Bhowmick P, Banerjee A, Chakrabarti S (2013). Antioxidant role of amyloid β protein in cell-free and biological systems: implication for the pathogenesis of Alzheimer disease. Free Radic Biol Med, 56: 184-92.
http://dx.doi.org/10.1016/j.freeradbiomed.2012.09.036
[25] Oliveri C, Ocaranza MP, Campos X, Lavandero S, Jalil JE (2001). Angiotensin I-converting enzyme modulates neutral endopeptidase activity in the rat. Hypertension, 38: 650-54.
http://dx.doi.org/10.1161/hy0901s1.095011
[26] Pfaffl MW (2001). A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res, 29: 2002-07.
[27] Fukumoto H, Rosene DL, Moss MB, Raju S, Hyman BT, Irizarry MC (2004). Beta-secretase activity increases with aging in human, monkey, and mouse brain. Am J Pathol, 164: 719-25.
http://dx.doi.org/10.1016/S0002-9440(10)63159-8
[28] Coulson DT, Beyer N, Quinn JG, Brockbank S, Hellemans J, Irvine GB, Ravid R, Johnston JA (2010). BACE1 mRNA expression in Alzheimer's disease postmortem brain tissue. J Alzheimers Dis, 22: 1111-22.
[29] El-Amouri SS, Zhu H, Yu J, Gage FH, Verma IM, Kindy MS (2007). Neprilysin protects neurons against Abeta peptide toxicity. Brain Res, 1152: 191-200.
http://dx.doi.org/10.1016/j.brainres.2007.03.072
[30] Miller BC, Eckman EA, Sambamurti K, Dobbs N, Chow KM, Eckman CB, Hersh LB, Thiele DL (2003). Amyloid-beta peptide levels in brain are inversely correlated with insulysin activity levels in vivo. Proc Natl Acad Sci U S A, 100: 6221-26.
http://dx.doi.org/10.1073/pnas.1031520100
[31] Wang S, Wang R, Chen L, Bennett DA, Dickson DW, Wang DS (2010). Expression and functional profiling of neprilysin, insulin-degrading enzyme, and endothelin-converting enzyme in prospectively studied elderly and Alzheimer's brain. J Neurochem, 115: 47-57.
http://dx.doi.org/10.1111/j.1471-4159.2010.06899.x
[32] Stockburger C, Gold VA, Pallas T, Kolesova N, Miano D, Leuner K, Müller WE (2014). A cell model for the initial phase of sporadic Alzheimer's disease. J Alzheimers Dis, 42: 395-411.
[33] Bobba A, Petragallo VA, Marra E, Atlante A (2010). Alzheimer's proteins, oxidative stress, and mitochondrial dysfunction interplay in a neuronal model of Alzheimer's disease. Int J Alzheimers Dis, 2010: 1-11.
[34] Yoon HS, Moon SC, Kim ND, Park BS, Jeong MH, Yoo YH (2000). Genistein induces apoptosis of RPE-J cells by opening mitochondrial PTP. Biochem Biophys Res Commun, 276: 151-56.
http://dx.doi.org/10.1006/bbrc.2000.3445
[35] Salvi M, Brunati AM, Clari G, Toninello A (2002). Interaction of genistein with the mitochondrial electron transport chain results in opening of the membrane transition pore. Biochim Biophys Acta, 1556: 187-96.
http://dx.doi.org/10.1016/S0005-2728(02)00361-4
[36] Xu J, Shi C, Li Q, Lam WP, Wai MS, Yew DT (2007). Effects of beta-amyloid peptide and estrogen on platelet mitochondrial function of Sprague-Dawley rats. Platelets, 18: 460-68.
http://dx.doi.org/10.1080/09537100701206808
[37] Joshi JV, Vaidya RA, Pandey SN, Agashe S, Chandrasekharan S, Menon SK, Vaidya AD (2007). Plasma levels of genistein following a single dose of soy extract capsule in Indian women. Indian J Med Res, 125: 534-41.
[38] Klein CB, King AA (2007). Genistein genotoxicity: critical considerations of in vitro exposure dose. Toxicol Appl Pharmacol, 224: 1-11.
http://dx.doi.org/10.1016/j.taap.2007.06.022
[39] Wiseman H, Casey K, Bowey EA, Duffy R, Davies M, Rowland IR, Lloyd AS, Murray A, Thompson R, Clarke DB (2004). Influence of 10 wk of soy consumption on plasma concentrations and excretion of isoflavonoids and on gut microflora metabolism in healthy adults. Am J Clin Nutr, 80: 692-99.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
Copyright © 2014 Aging and Disease, All Rights Reserved.
Address: Aging and Disease Editorial Office 3400 Camp Bowie Boulevard Fort Worth, TX76106 USA
Fax: (817) 735-0408 E-mail: editorial@aginganddisease.org
Powered by Beijing Magtech Co. Ltd