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Anti-Cancer Agents in Medicinal Chemistry

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ISSN (Print): 1871-5206
ISSN (Online): 1875-5992

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Research Article

Anticancer Effect of Amygdalin (Vitamin B-17) on Hepatocellular Carcinoma Cell Line (HepG2) in the Presence and Absence of Zinc

Author(s): Mohamed A. El-Desouky, Abdelgawad A. Fahmi*, Ibrahim Y. Abdelkader and Karima M. Nasraldin

Volume 20, Issue 4, 2020

Page: [486 - 494] Pages: 9

DOI: 10.2174/1871520620666200120095525

Price: $65

Abstract

Background: Amygdalin (Vitamin B-17) is a naturally occurring vitamin found in the seeds of the fruits of Prunus Rosacea family including apricot, bitter almond, cherry, and peach.

Objective: The purpose of this study was to examine the effect of amygdalin with and without zinc on hepatocellular carcinoma (HepG2) cell line.

Methods: MTT assay was used to evaluate the cytotoxicity of amygdalin without zinc, amygdalin + 20μmol zinc, and amygdalin + 800μmol zinc on HepG2 cell lines. The cell cycle distribution assay was determined by flow cytometry. Apoptosis was confirmed by Annexin V-FITC/PI staining assay. Moreover, the pathway of apoptosis was determined by the percentage of change in the mean levels of P53, Bcl2, Bax, cytochrome c, and caspase-3.

Results: Amygdalin without zinc showed strong anti-HepG2 activity. Furthermore, HepG2 cell lines treatment with amygdalin + 20μmol zinc and amygdalin + 800μmol zinc showed a highly significant apoptotic effect than the effect of amygdalin without zinc. Amygdalin treatment induced cell cycle arrest at G2/M and increased the levels of P53, Bax, cytochrome c, and caspase-3 significantly, while it decreased the level of anti-apoptotic Bcl2.

Conclusion: Amygdalin is a natural anti-cancer agent, which can be used for the treatment of hepatocellular carcinoma. It promotes apoptosis via the intrinsic cell death pathway (the mitochondria-initiated pathway) and cell cycle arrest at G/M. The potency of amygdalin in HepG2 treatment increased significantly by the addition of zinc.

Keywords: Amygdalin, vitamin B-17, hepatocellular carcinoma, anti-cancer, Bcl2, caspase-3, P53, zinc.

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[1]
Tang, A.; Hallouch, O.; Chernyak, V.; Kamaya, A.; Sirlin, C.B. Epidemiology of hepatocellular carcinoma: target population for surveillance and diagnosis. Abdom. Radiol. (N.Y.), 2018, 43(1), 13-25.
[http://dx.doi.org/10.1007/s00261-017-1209-1] [PMID: 28647765]
[2]
Song, M.J.; Bae, S.H. Newer treatments for advanced hepatocellular carcinoma. Korean J. Intern. Med. (Korean. Assoc. Intern. Med.), 2014, 29(2), 149-155.
[http://dx.doi.org/10.3904/kjim.2014.29.2.149] [PMID: 24648795]
[3]
Hwang, H-J.; Lee, H-J.; Kim, C-J.; Shim, I.; Hahm, D-H. Inhibitory effect of amygdalin on lipopolysaccharide-inducible TNF-alpha and IL-1beta mRNA expression and carrageenan-induced rat arthritis. J. Microbiol. Biotechnol., 2008, 18(10), 1641-1647.
[PMID: 18955812]
[4]
Ioannis, P.; Anastasis, S.; Andreas, Y. Tripterygium wilfordii extract (Triptolide) and amygdalin promotes cell death in cancer cells: True or a myth. Am. J. Cancer Prev., 2015, 3(4), 77-83.
[http://dx.doi.org/10.12691/ajcp-3-4-3]
[5]
Enculescu, M. Vitamin B 17/Laetrile/Amygdalin (a Review). Bull. Univ. Agric. Sci. Vet. Med. Cluj-Napoca Anim. Sci. Biotechnol., 2009, 66(11), 20-25.
[6]
Franklin, R.B.; Costello, L.C. The important role of the apoptotic effects of zinc in the development of cancers. J. Cell. Biochem., 2009, 106(5), 750-757.
[http://dx.doi.org/10.1002/jcb.22049] [PMID: 19160419]
[7]
http://www.novodalin.com/
[8]
https://www.tjsupply.com/vitamin-b17-injections.html
[9]
https://www.sigmaaldrich.com/MSDS/MSDS/DisplayMS
[10]
Knowles, B.B.; Howe, C.C.; Aden, D.P. Human hepatocellular carcinoma cell lines secrete the major plasma proteins and hepatitis B surface antigen. Science, 1980, 209(4455), 497-499.http://dx.doi.org///doi.org/10.1126/science.6248960
[11]
Sylvester, P.W. Optimization of the tetrazolium dye (MTT) colorimetric assay for cellular growth and viability. Methods Mol. Biol., 2011, 716, 157-168.
[http://dx.doi.org/10.1007/978-1-61779-012-6_9] [PMID: 21318905]
[12]
Pozarowski, P.; Darzynkiewicz, Z. Analysis of cell cycle by flow cytometry. Methods Mol. Biol., 2004, 281, 301-311.
[http://dx.doi.org/10.1385/1-59259-811-0:301] [PMID: 15220539]
[13]
Zhao, G.; Han, X.; Cheng, W.; Ni, J.; Zhang, Y.; Lin, J.; Song, Z. Apigenin inhibits proliferation and invasion, and induces apoptosis and cell cycle arrest in human melanoma cells. Oncol. Rep., 2017, 37(4), 2277-2285.
[http://dx.doi.org/10.3892/or.2017.5450] [PMID: 28260058]
[14]
Balmer, M.T.; Katz, R.D.; Liao, S.; Goodwine, J.S.; Gal, S. Doxorubicin and 5-fluorouracil induced accumulation and transcriptional activity of p53 are independent of the phosphorylation at serine 15 in MCF-7 breast cancer cells. Cancer Biol. Ther., 2014, 15(8), 1000-1012.
[http://dx.doi.org/10.4161/cbt.29112] [PMID: 24801380]
[15]
Adhami, V.M.; Aziz, M.H.; Mukhtar, H.; Ahmad, N. Activation of prodeath Bcl-2 family proteins and mitochondrial apoptosis pathway by sanguinarine in immortalized human HaCaT keratinocytes. Clin. Cancer Res., 2003, 9(8), 3176-3182.
[PMID: 12912970]
[16]
Oguchi, T.; Ono, R.; Tsuji, M.; Shozawa, H.; Somei, M.; Inagaki, M.; Mori, Y.; Yasumoto, T.; Ono, K.; Kiuchi, Y. Cilostazol suppresses Aβ-induced neurotoxicity in SH-SY5Y cells through inhibition of oxidative stress and MAPK signaling pathway. Front. Aging Neurosci., 2017, 9, 337.
[http://dx.doi.org/10.3389/fnagi.2017.00337] [PMID: 29089887]
[17]
Manickam, P.; Kaushik, A.; Karunakaran, C.; Bhansali, S. Recent advances in cytochrome c biosensing technologies. Biosens. Bioelectron., 2017, 87, 654-668.
[http://dx.doi.org/10.1016/j.bios.2016.09.013] [PMID: 27619529]
[18]
Schroeter, H.; Spencer, J.P.; Rice-Evans, C.; Williams, R.J. Flavonoids protect neurons from oxidized low-density-lipoprotein-induced apoptosis involving c-Jun N-terminal kinase (JNK), c-Jun and caspase-3. Biochem. J., 2001, 358(Pt 3), 547-557.
[http://dx.doi.org/10.1042/bj3580547] [PMID: 11535118]
[19]
Nahrstedt, A.; Sattar, E.A.; El-Zalabani, M.H.S. Amygdalin acyl derivatives, cyanogenic glycosides from the seeds of Merremia dissecta. Phytochemistry, 1990, 29(4), 1179-1181.
[http://dx.doi.org/10.1016/0031-9422(90)85424-E]
[20]
Zhou, C.; Qian, L.; Ma, H.; Yu, X.; Zhang, Y.; Qu, W.; Zhang, X.; Xia, W. Enhancement of amygdalin activated with β-D-glucosidase on HepG2 cells proliferation and apoptosis. Carbohydr. Polym., 2012, 90(1), 516-523.
[http://dx.doi.org/10.1016/j.carbpol.2012.05.073] [PMID: 24751072]
[21]
Wang, G-W.; Lv, C.; Shi, Z-R.; Zeng, R-T.; Dong, X-Y.; Zhang, W-D.; Liu, R-H.; Shan, L.; Shen, Y-H. Abieslactone induces cell cycle arrest and apoptosis in human hepatocellular carcinomas through the mitochondrial pathway and the generation of reactive oxygen species. PLoS One, 2014, 9(12) e115151
[http://dx.doi.org/10.1371/journal.pone.0115151] [PMID: 25502685]
[22]
Guo, J-R.; Chen, Q-Q.; Lam, C.W.; Zhang, W. Effects of karanjin on cell cycle arrest and apoptosis in human A549, HepG2 and HL-60 cancer cells. Biol. Res., 2015, 48, 40.
[http://dx.doi.org/10.1186/s40659-015-0031-x] [PMID: 26209237]
[23]
Gao, F.; Ma, N.; Zhou, H.; Wang, Q.; Zhang, H.; Wang, P.; Hou, H.; Wen, H.; Li, L. Zinc oxide nanoparticles-induced epigenetic change and G2/M arrest are associated with apoptosis in human epidermal keratinocytes. Int. J. Nanomedicine, 2016, 11, 3859-3874.
[http://dx.doi.org/10.2147/IJN.S107021] [PMID: 27570453]
[24]
Green, D.R.; Kroemer, G. Cytoplasmic functions of the tumour suppressor p53. Nature, 2009, 458(7242), 1127-1130.
[http://dx.doi.org/10.1038/nature07986] [PMID: 19407794]
[25]
Amirah, A. Preventive role of amygdalin in squamous cell carcinoma induced in hamsters. Int. J. Pharm. Sci. Med., 2017, 2, 8-14.
[26]
Provinciali, M.; Pierpaoli, E.; Bartozzi, B.; Bernardini, G. Zinc induces apoptosis of human melanoma cells, increasing reactive oxygen species, p53 and FAS ligand. Anticancer Res., 2015, 35(10), 5309-5316.
[PMID: 26408691]
[27]
Dhawan, D.K.; Chadha, V.D. Zinc: a promising agent in dietary chemoprevention of cancer. Indian J. Med. Res., 2010, 132, 676-682.
[28]
Duffy, M.J.; Synnott, N.C.; Crown, J. Mutant p53 as a target for cancer treatment. Eur. J. Cancer, 2017, 83, 258-265.
[http://dx.doi.org/10.1016/j.ejca.2017.06.023] [PMID: 28756138]
[29]
Lehman-McKeeman, L.D. Mechanisms of Toxicity.Casarett & Doull’s Toxicology: The Basic Science of Poisons, 9th ed; Klaassen, C.D., Ed.; McGraw-Hill Education: New York, NY, 2019.
[30]
Reddy, L.; Odhav, B.; Bhoola, K. Aflatoxin B1-induced toxicity in HepG2 cells inhibited by carotenoids: morphology, apoptosis and DNA damage. Biol. Chem., 2006, 387(1), 87-93.
[http://dx.doi.org/10.1515/BC.2006.012] [PMID: 16497168]
[31]
Jumaa, A.H.; Al-Uboody, W.S.H.; Hady, A.M. Esomeprazole and amygdalin combination cytotoxic effect on human cervical cancer cell line (hela cancer cell line). J. Pharm. Sci. Res., 2018, 10(9), 2236-2241.
[32]
Babula, P.; Kohoutkova, V.; Opatrilova, R.; Dankova, I.; Masarik, M.; Kizek, R. Pharmaceutical importance of zinc and metallothionein in cell signaling. Pharmaceut. Anal., 2010, 28, 18-21.
[33]
Moon, J-Y.; Kim, S-W.; Yun, G-M.; Lee, H-S.; Kim, Y-D.; Jeong, G-J.; Ullah, I.; Rho, G-J.; Jeon, B-G. Inhibition of cell growth and down-regulation of telomerase activity by amygdalin in human cancer cell lines. Anim. Cells Syst., 2015, 19(5), 295-304.
[http://dx.doi.org/10.1080/19768354.2015.1060261]
[34]
Feng, P.; Li, T.; Guan, Z.; Franklin, R.B.; Costello, L.C. The involvement of Bax in zinc-induced mitochondrial apoptogenesis in malignant prostate cells. Mol. Cancer, 2008, 7, 25.
[http://dx.doi.org/10.1186/1476-4598-7-25] [PMID: 18331646]
[35]
Cui, A.; Wei, C.; Yang, Q.; Li, F. Amygdalin induces apoptosis in human cervical cancer cell line HeLa cells. Immunopharmacol. Immunotoxicol., 2013, 35(1), 43-51.http://dx.doi.org/https://doi.org/10.3109/08923973.2012.738688
[36]
Chang, H.K.; Shin, M.S.; Yang, H.Y.; Lee, J.W.; Kim, Y.S.; Le, M.H.; Kim, J.; Kim, K.H.; Kim, C.J. Amygdalin induces apoptosis through regulation of Bax and Bcl-2 expressions in human DU145 and LNCaP prostate cancer cells. Biol. Pharm. Bull., 2006, 29(8), 1597-1602.
[37]
Feng, P.; Li, T.; Guan, Z.; Franklin, R.B.; Costello, L.C. The involvement of Bax in zinc-induced mitochondrial apoptogenesis in malignant prostate cells. Mol. Cancer, 2008, 7, 25.
[http://dx.doi.org/10.1186/1476-4598-7-25]
[38]
Sireesha, D.; Reddy, B.S.; Reginald, B.A.; Samatha, M.; Kamal, F. Effect of amygdalin on oral cancer cell line: An in vitro study. J. Oral Maxillofac. Pathol., 2019, 23(1), 104-107.
[http://dx.doi.org/10.4103/jomfp.JOMFP_281_18] [PMID: 31110425]
[39]
Eron, S.J.; MacPherson, D.J.; Dagbay, K.B.; Hardy, J.A. Multiple mechanisms of zinc-mediated inhibition for the apoptotic caspases-3, -6, -7, and -8. ACS Chem. Biol., 2018, 13(5), 1279-1290.
[http://dx.doi.org/10.1021/acschembio.8b00064] [PMID: 29364645]
[40]
Velázquez-Delgado, E.M.; Hardy, J.A. Zinc-mediated allosteric inhibition of caspase-6. J. Biol. Chem., 2012, 287(43), 36000-36011.
[http://dx.doi.org/10.1074/jbc.M112.397752] [PMID: 22891250]
[41]
Schrantz, N.; Auffredou, M.T.; Bourgeade, M.F.; Besnault, L.; Leca, G.; Vazquez, A. Zinc-mediated regulation of caspases activity: dose-dependent inhibition or activation of caspase-3 in the human Burkitt lymphoma B cells (Ramos). Cell Death Differ., 2001, 8(2), 152-161.
[http://dx.doi.org/10.1038/sj.cdd.4400772] [PMID: 11313717]
[42]
Autret, A.; Martin, S.J. Emerging role for members of the Bcl-2 family in mitochondrial morphogenesis. Mol. Cell, 2009, 36(3), 355-363.
[http://dx.doi.org/10.1016/j.molcel.2009.10.011] [PMID: 19917245]
[43]
Chang, H-K.; Shin, M-S.; Yang, H-Y.; Lee, J-W.; Kim, Y-S.; Lee, M-H.; Kim, J.; Kim, K-H.; Kim, C-J. Amygdalin induces apoptosis through regulation of Bax and Bcl-2 expressions in human DU145 and LNCaP prostate cancer cells. Biol. Pharm. Bull., 2006, 29(8), 1597-1602.
[http://dx.doi.org/10.1248/bpb.29.1597] [PMID: 16880611]
[44]
Song, Z.; Xu, X. Advanced research on anti-tumor effects of amygdalin. J. Cancer Res. Ther., 2014, 10(Suppl. 1), 3-7.
[http://dx.doi.org/10.4103/0973-1482.139743] [PMID: 25207888]
[45]
Liang, J.Y.; Feng, P.; Li, T.L.; Guan, Z.X.; Zou, J.; Franklin, R.; Costello, L.C. Zinc induces mitochondria apoptogenesis in prostate cells. Mol. Urol., 2004, 4(1), 31-36.
[46]
Fukuda, T.; Ito, H.; Mukainaka, T.; Tokuda, H.; Nishino, H.; Yoshida, T. Anti-tumor promoting effect of glycosides from Prunus persica seeds. Biol. Pharm. Bull., 2003, 26(2), 271-273.
[http://dx.doi.org/10.1248/bpb.26.271] [PMID: 12576693]
[47]
Lee, H.M.; Moon, A. Amygdalin regulates apoptosis and adhesion in Hs578T triple-negative breast cancer cells. Biol. Ther. (Seoul), 2016, 24(1), 62-66.http://dx.doi.org/ https://doi.org/10.4062/biomolther.2015.172
[48]
Chen, Y.; Ma, J.; Wang, F.; Hu, J.; Cui, A.; Wei, C.; Yang, Q.; Li, F. Amygdalin induces apoptosis in human cervical cancer cell line HeLa cells. Immunopharmacol. Immunotoxicol., 2013, 35(1), 43-51.
[http://dx.doi.org/10.3109/08923973.2012.738688] [PMID: 23137229]
[49]
Juengel, E.; Thomas, A.; Rutz, J.; Makarevic, J.; Tsaur, I.; Nelson, K.; Haferkamp, A.; Blaheta, R.A. Amygdalin inhibits the growth of renal cell carcinoma cells in vitro. Int. J. Mol. Med., 2016, 37(2), 526-532.
[http://dx.doi.org/10.3892/ijmm.2015.2439] [PMID: 26709398]
[50]
Makarević, J.; Rutz, J.; Juengel, E.; Kaulfuss, S.; Tsaur, I.; Nelson, K.; Pfitzenmaier, J.; Haferkamp, A.; Blaheta, R.A. Amygdalin influences bladder cancer cell adhesion and invasion in vitro. PLoS One, 2014, 9(10) e110244
[http://dx.doi.org/10.1371/journal.pone.0110244] [PMID: 25333694]
[51]
Choi, S.; Cui, C.; Luo, Y.; Kim, S-H.; Ko, J-K.; Huo, X.; Ma, J.; Fu, L-W.; Souza, R.F.; Korichneva, I.; Pan, Z. Selective inhibitory effects of zinc on cell proliferation in esophageal squamous cell carcinoma through Orai1. FASEB J., 2018, 32(1), 404-416.
[http://dx.doi.org/10.1096/fj.201700227RRR] [PMID: 28928244]

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