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ISSN (Print): 2212-7984
ISSN (Online): 1876-1429

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Research Article

Evaluation of the Cytotoxicity and Apoptotic Induction in Human Liver Cell Lines Exposed to Three Food Additives

Author(s): Ingy M. El-Hefny*, Neima K. Al Senosy, Walaa G. Hozayen, Amr E. Ahmed, Ayman Diab and Wesam T. Basal

Volume 11, Issue 3, 2020

Page: [193 - 201] Pages: 9

DOI: 10.2174/2212798411666200217124630

Abstract

Background: Rapid lifestyle, especially among people living in urban areas, has led to increasing reliance on the processed food market. Unfortunately, harmful effects caused by the excessive use of food additives in such type of industry are often neglected.

Objective: This proposal investigates in vitro cytotoxic and apoptotic effects of three food preservatives commonly consumed in daily meals; sodium sulphite, boric acid, and benzoic acid.

Methods: The effect of the three preservatives on cell viability was tested on two different cell lines; normal liver cell line THLE2 and human hepatocellular carcinoma cancer cell line HepG2 using MTT assay. Cell cycle arrest was measured using flow cytometry by propidium iodide. Measurement of expression levels of two central genes, p53 and bcl-2 that play key roles in cell cycle and apoptosis was carried out in HepG2 cells using real time-PCR.

Results: Although the effect was more significantly realized in the HepG2 cell line, the viability of both cell lines was decreased by all of the three tested compounds. Flow cytometric analysis of HepG2 cells treated with sodium sulphite, boric acid, and benzoic acid has revealed an increase in G2/M phase cell cycle arrest. In Sodium sulphite and boric acid-treated cells, expression levels of p53 were up-regulated, while that of the Bcl2 was significantly down-regulated. On the other hand, Benzoic acid has shown an anti-apoptotic feature based on the increased expression levels of Bcl-2 in treated cells.

Conclusion: In conclusion, all of the tested compounds have decreased the cell line viability and induced both cell cycle arrest and apoptotic events indicating their high potential of being cytotoxic and genotoxic materials.

Keywords: Cytotoxic, genotoxic, cell cycle arrest, real-time PCR, cell lines, food additives.

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[1]
Rekha K, Dharman AK. Mitotic aberrations induced by sodium benzoate: a food additive in Allium cepa L. Plant Arch 2011; 11(2): 945-7.
[2]
Tuormaa TE. The adverse effects of food additives on health: a review of the literature with a special emphasis on childhood hyperactivity. J Orthomol Med 1994; 9: 225-43.
[3]
Dreisig K, Taxvig C, Birkhøj Kjærstad M, Nellemann C, Hass U, Vinggaard AM. Predictive value of cell assays for developmental toxicity and embryotoxicity of conazole fungicides. ALTEX 2013; 30(3): 319-30.
[http://dx.doi.org/10.14573/altex.2013.3.319] [PMID: 23861077]
[4]
Das A, Mukherjee A. Genotoxicity testing of the food colours amaranth and tartrazineInt J Hum Genet 2004; 4(4): 277-80.
[http://dx.doi.org/10.1080/09723757.2004.11885906]
[5]
Growther L, Parimala R, Karthiga G, Vimalin J, Kalimuthu K, Sangeetha K B. Food Additives and Their Mutagenicity Int J Nutrition Wellness 2008; 7(2)
[6]
Aeschbacher HU. Genetic toxicology of food products. In: Mendelsohn ML, Albertini RJ, Eds Mutation and the Environment Part E: Environmental Genotoxicity, Risk and modulation. New York: Wiley-Liss 1990; pp. 117-26.
[7]
Wakabayashi K. Identification of food mutagens.In: Mendelsohn ML, Albertini RJ, Eds Mutation and the Environment Part E: Environmental Genotoxicity, Risk and modulation. New York: Wiley-Liss 1990; pp. 107-16.
[8]
Tanaka R. Inhibitory effects of xanthone on paraquat- and NaNO(2)-induced genotoxicity in cultured cells. J Toxicol Sci 2007; 32(5): 571-4.
[http://dx.doi.org/10.2131/jts.32.571] [PMID: 18198487]
[9]
Türkoğlu S. Genotoxicity of five food preservatives tested on root tips of Allium cepa L Mutat Res 2007; 626(1-2): 4-14.
[http://dx.doi.org/10.1016/j.mrgentox.2006.07.006] [PMID: 17005441]
[10]
Demir E, Kocaoğlu S, Kaya B. Genotoxicity testing of four benzyl derivatives in the Drosophila wing spot test. Food Chem Toxicol 2008; 46(3): 1034-41.
[http://dx.doi.org/10.1016/j.fct.2007.10.035] [PMID: 18068884]
[11]
Auerbach C. Mutation research - Problems, Results and Perspectives. Chapman and Hall London 1976.
[http://dx.doi.org/10.1007/978-1-4899-3103-0]
[12]
Al-Tai Protective Effect of pomegranate molasses (PM) against genotoxicity induced by Benzoic acid (E-210) in human lymphocytes in vitro. Nat Sci 2014; 12(11): 13-6.
[13]
Barranco WT, Eckhert CD. Boric acid inhibits human prostate cancer cell proliferationCancer Lett 2004; 216(1): 21-9.
[http://dx.doi.org/10.1016/j.canlet.2004.06.001] [PMID: 15500945]
[14]
Wei Y, Yuan FJ, Zhou WB, et al. Borax-induced apoptosis in HepG2 cells involves p53, Bcl-2, and Bax Genet Mol Res 2016; 5(2)
[http://dx.doi.org/10.4238/gmr.15028300] [PMID: 27420953]
[15]
Ekwall B. Correlation between cytotoxicity in vitro and LD50-valuesActa Pharmacol Toxicol (Copenh) 1983; 52(Suppl. 2): 80-99.
[http://dx.doi.org/10.1111/j.1600-0773.1983.tb02685.x] [PMID: 6880789]
[16]
Yamashoji S, Isshiki K. Rapid detection of cytotoxicity of food additives and contaminants by a novel cytotoxicity test, menadione-catalyzed H(2)O (2) production assay. Cytotechnology 2001; 37(3): 171-8.
[http://dx.doi.org/10.1023/A:1020580818979] [PMID: 19002920]
[17]
Gilbert DF, Friedrich O. Cell viability assays: methods and protocols. 1st ed. Humana Press: New Jersey 2017..
[18]
Berridge MV, Herst PM, Tan AS. Tetrazolium dyes as tools in cell biology: New insights into their cellular reduction. In: El-Gewely MR, Ed Biotechnology annual review. Elsevier 2005; pp. 127-52.
[http://dx.doi.org/10.1016/S1387-2656(05)11004-7]
[19]
Riss TL, Moravec RA, Niles AL, et al. Cell viability assays. In: Sittampalam GS, Coussens NP, Nelson H, et al Eds Assay guidance manual. Bethesda, MD: Eli Lilly & Company and the National Center for Advancing Translational Sciences 2013.
[20]
Lewis CW, Smith JE, Anderson JG, Freshney RI. Increased cytotoxicity of food-borne mycotoxins toward human cell lines in vitro via enhanced cytochrome p450 expression using the MTT bioassay. Mycopathologia 1999; 148(2): 97-102.
[http://dx.doi.org/10.1023/A:1007130923558] [PMID: 11189750]
[21]
Bonelli G, Sacchi MC, Barbiero G, et al. Apoptosis of L929 cells by etoposide: a quantitative and kinetic approach. Exp Cell Res 1996; 228(2): 292-305.
[http://dx.doi.org/10.1006/excr.1996.0329] [PMID: 8912723]
[22]
Wu ZZ, Chien CM, Yang SH, et al. Induction of G2/M phase arrest and apoptosis by a novel enediyne derivative, THDA, in chronic myeloid leukemia (K562) cells. Mol Cell Biochem 2006; 292(1-2): 99-105.
[http://dx.doi.org/10.1007/s11010-006-9222-7] [PMID: 16733797]
[23]
Crissman HA, Hirons GT. Staining of DNA in live and fixed cellsMethods Cell Biol 1994; 41: 195-209.
[http://dx.doi.org/10.1016/S0091-679X(08)61718-5] [PMID: 7532262]
[24]
Darzynkiewicz Z, Gong J, Juan G, Ardelt B, Traganos F. Cytometry of cyclin proteinsCytometry 1996; 25(1): 1-13.
[http://dx.doi.org/10.1002/(SICI)1097-0320(19960901)25:1<1::AID-CYTO1>3.0.CO;2-N] [PMID: 8875049]
[25]
Larsen JK, Landberg G, Roos G. Detection of proliferating cell nuclear antigen Methods Cell Biol 2001; 63: 419-31.
[http://dx.doi.org/10.1016/S0091-679X(01)63023-1] [PMID: 11060852]
[26]
Esmaeili-Mahani S, Falahi F, Yaghoobi MM. roapoptotic and antiproliferative effects of Thymus caramanicus on human breast cancer cell line (MCF-7) and its interaction with anticancer drug vincristine Evid Based Complement Alternat Med 2014.2014893247.
[http://dx.doi.org/10.1155/2014/893247] [PMID: 24812569]
[27]
Roshan S, Liu YY, Banafa A, et al. Fucoidan induces apoptosis of HepG2 cells by down-regulating p-Stat3. J Huazhong Univ Sci Technolog Med Sci 2014; 34(3): 330-6.
[http://dx.doi.org/10.1007/s11596-014-1278-0] [PMID: 24939294]
[28]
Kerr JF, Wyllie AH, Currie AR. Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer 1972; 26(4): 239-57.
[http://dx.doi.org/10.1038/bjc.1972.33] [PMID: 4561027]
[29]
Raffray M, Cohen GM. Apoptosis and necrosis in toxicology: a continuum or distinct modes of cell death? Pharmacol Ther 1997; 75(3): 153-77.
[http://dx.doi.org/10.1016/S0163-7258(97)00037-5] [PMID: 9504137]
[30]
Issaeva N, Bozko P, Enge M, et al. Small molecule RITA binds to p53, blocks p53-HDM-2 inter-action and activates p53 function in tumors. Nat Med 2004; 10(12): 1321-8.
[http://dx.doi.org/10.1038/nm1146]
[31]
Levine AJ. p53, the cellular gatekeeper for growth and division Cell 1997; 88(3): 323-1.
[http://dx.doi.org/10.1016/S0092-8674(00)81871-1] [PMID: 9039259]
[32]
Haupt S, Haupt Y. Manipulation of the tumor suppressor p53 for potentiating cancer therapy. Semin Cancer Biol 2004; 14(4): 244-52.
[http://dx.doi.org/10.1016/j.semcancer.2004.04.003] [PMID: 15219617]
[33]
Mirzayans R, Andrais B, Scott A, Murray D. New insights into p53 signaling and cancer cell response to DNA damage: implications for cancer therapy. J Biomed Biotechnol 2012; •••2012170325
[http://dx.doi.org/10.1155/2012/170325] [PMID: 22911014]
[34]
Dewson G, Kluck RM. Mechanisms by which Bak and Bax permeabilise mitochondria during apoptosis. J Cell Sci 2009; 122(Pt 16): 2801-8.
[http://dx.doi.org/10.1242/jcs.038166] [PMID: 19795525]
[35]
Wu M, Zhang H, Hu J, et al. Isoalantolactone inhibits UM-SCC-10A cell growth via cell cycle arrest and apoptosis induction. PLoS One 2013; 8(9)e76000
[http://dx.doi.org/10.1371/journal.pone.0076000] [PMID: 24098753]
[36]
Chen Q, Lesnefsky EJ. Blockade of electron transport during ischemia preserves bcl-2 and inhibits opening of the mitochondrial permeability transition pore. FEBS Lett 2011; 585(6): 921-6.
[http://dx.doi.org/10.1016/j.febslet.2011.02.029] [PMID: 21354418]
[37]
See AS, Salleh AB, Bakar FA, Yusof NA, Abdulamir AS, Heng LY. Risk and health effect of boric acidAm J Appl Sci 2010; 7: 620-7.
[http://dx.doi.org/10.3844/ajassp.2010.620.627]
[38]
Litovitz TL, Klein-Schwartz W, Oderda GM, Schmitz BF. Clinical manifestations of toxicity in a series of 784 boric acid ingestions. Am J Emerg Med 1988; 6(3): 209-13.
[http://dx.doi.org/10.1016/0735-6757(88)90001-0] [PMID: 3370093]
[39]
Canturk Z, Tunali Y, Korkmaz S, Gulbaş Z. Cytotoxic and apoptotic effects of boron compounds on leukemia cell line. Cytotechnology 2016; 68(1): 87-93.
[http://dx.doi.org/10.1007/s10616-014-9755-7] [PMID: 25159521]
[40]
Murmu N, Ghosh P, Gomes A, et al. Antineoplastic effect of new boron compounds against leukemic cell lines and cells from leukemic patients. J Exp Clin Cancer Res 2002; 21(3): 351-6.
[PMID: 12385577]
[41]
Gallardo-Williams MT, Maronpot RR, Wine RN, Brunssen SH, Chapin RE. Inhibition of the enzymatic activity of prostate-specific antigen by boric acid and 3-nitrophenyl boronic acid. Prostate 2003; 54(1): 44-9.
[http://dx.doi.org/10.1002/pros.10166] [PMID: 12481254]
[42]
Yilmaz B, Karabay AZ. Food additive sodium benzoate (NaB) activates NFκB and induces apoptosis in HCT116 Cells. Molecules 2018; 23(4): 723.
[http://dx.doi.org/10.3390/molecules23040723] [PMID: 29565269]
[43]
Park H, Park EH, Yun H. Sodium Benzoate-mediated cytotoxicitymammalian cells. J Food Biochem 2011; 35(4): 1034-46.
[44]
Yadav A, Kumar A, Das M, Tripathi A. Sodium benzoate, a food preservative, affects the functional and activation status of splenocytes at non cytotoxic dose. Food Chem Toxicol 2016; 88: 40-7.
[http://dx.doi.org/10.1016/j.fct.2015.12.016] [PMID: 26706697]
[45]
Spindola DG, Hinsberger A, Antunes VM, Michelin L, Claudia B, Carlos RO. In vitro cytotoxicity of chemical preservatives on human fibroblast cells. Braz J Pharm Sci 2018; 54(1)
[http://dx.doi.org/10.1590/s2175-97902018000100031]
[46]
Sleiman RJ, Stewart BW. Early caspase activation in leukemic cells subject to etoposide-induced G2-M arrest: evidence of commitment to apoptosis rather than mitotic cell death. Clin Cancer Res 2000; 6(9): 3756-65.
[PMID: 10999770]
[47]
Chung TW, Lin SC, Su JH, Chen YK, Lin CC, Chan HL. Sinularin induces DNA damage, G2/M phase arrest, and apoptosis in human hepatocellular carcinoma cells. BMC Complement Altern Med 2017; 17(1): 62.
[http://dx.doi.org/10.1186/s12906-017-1583-9] [PMID: 28103869]
[48]
Neima K. The phathalate DBP-induced cytotoxicity and apoptosis via gene expression of p53, Bcl2 and Bax in tumor cell lines. Egypt J Genet Cytol 2018; 47(1): 45-56.
[49]
Aardema MJ, MacGregor JT. Toxicology and genetic toxicology in the new era of “toxicogenomics”: impact of “-omics” technologies. Mutat Res 2002; 499(1): 13-25.
[http://dx.doi.org/10.1016/S0027-5107(01)00292-5] [PMID: 11804602]
[50]
Strasser A, Harris AW, Jacks T, Cory S. DNA damage can induce apoptosis in proliferating lymphoid cells via p53-independent mechanisms inhibitable by Bcl-2. Cell 1994; 79(2): 329-39.
[http://dx.doi.org/10.1016/0092-8674(94)90201-1] [PMID: 7954799]
[51]
Kirsch-Volders M, Vanhauwaert A, Eichenlaub-Ritter U, Decordier I. Indirect mechanisms of genotoxicity. Toxicol Lett 2003; 140-141: 63-74.
[http://dx.doi.org/10.1016/S0378-4274(02)00498-8] [PMID: 12676452]
[52]
Choi JH, Ahn KS, Kim J, Hong YS. Enhanced induction of Bax gene expression in H460 and H1299 cells with the combined treatment of cisplatin and adenovirus mediated wt-p53 gene transfer. Exp Mol Med 2000; 32(1): 23-8.
[http://dx.doi.org/10.1038/emm.2000.5] [PMID: 10762058]
[53]
Polager S, Ginsberg D. p53 and E2f: partners in life and death Nat Rev Cancer 2009; 910; : 738-48.
[http://dx.doi.org/10.1038/nrc2718] [PMID: 19776743]
[54]
Leu JI, Dumont P, Hafey M, Murphy ME, George DL. Mitochondrial p53 activates Bak and causes disruption of a Bak-Mcl1 complex. Nat Cell Biol 2004; 6(5): 443-50.
[http://dx.doi.org/10.1038/ncb1123] [PMID: 15077116]
[55]
Li F, Srinivasan A, Wang Y, Armstrong RC, Tomaselli KJ, Fritz LC. Cell-specific induction of apoptosis by microinjection of cytochrome c. Bcl-xL has activity independent of cytochrome c release. J Biol Chem 1997; 272(48): 30299-305.
[http://dx.doi.org/10.1074/jbc.272.48.30299] [PMID: 9374516]
[56]
Raposa B, Pónusz R, Gerencsér G, et al. Food additives: sodium benzoate, potassium sorbate, azorubine, and tartrazine modify the expression of NFκB, GADD45α, and MAPK8 genes. Physiol Int 2016; 103(3): 334-43.
[http://dx.doi.org/10.1556/2060.103.2016.3.6] [PMID: 28229641]
[57]
GAO Y. Effect of food azo dye atrazine on learning and memory functions in mice and rats, and the possible mechanisms involved. J Food Sci 2011; 76: 125-9.
[http://dx.doi.org/10.1111/j.1750-3841.2011.02267.x]

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