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Clinical Cancer Drugs

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ISSN (Print): 2212-697X
ISSN (Online): 2212-6988

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Research Article

Melatonin Induces Apoptosis and Inhibits the Proliferation of Cancer Cells via Reactive Oxygen Species-mediated MAPK and mTOR Pathways

Author(s): Reena Kasi, Pei Ling Yeo, Ng. Khuen Yen, Rhun Yian Koh, Gnanajothy Ponnudurai, Yee Lian Tiong and Soi Moi Chye*

Volume 7, Issue 1, 2020

Page: [44 - 56] Pages: 13

DOI: 10.2174/2212697X06666191116151114

Abstract

Background: Recent human and animal studies have demonstrated the oncostatic properties of N-acetyl-5-methoxytryptamine (melatonin) in different types of cancer. However, in few cancer cell lines including colorectal cancer cell line (HT-29), acute T cell leukemia cell line (JURKAT) and cervical cancer cell line (HeLa), precise oncostatic mechanism induced by melatonin is yet to be described.

Objectives: The aim of this study is to investigate the effects of melatonin in HT-29, JURKAT and HeLa cells and to determine the underlying molecular mechanism.

Methods: Cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay while cell cycle, apoptosis and membrane potential were analysed by flow cytometry. Reactive oxygen species (ROS) was detected by 2',7'-dichlorofluorescein diacetate (DCFH-DA) staining. Protein expressions were determined by Western blot.

Results: Our results showed that melatonin suppressed cell proliferation, increased the number of sub G1 hypodiploid cells and cell cycle arrest in HT-29, JURKAT and HeLa cells. Besides, melatonin also induced early and late apoptosis, although there were marked variations in responses between different cell lines (sensitivity; HeLa > HT-29 >JURKAT). Apart from that, staining with DCHF-DA demonstrated ROS production that was induced in a dose-dependent manner in HeLa, HT-29 and JURKAT cells. Moreover, the apoptotic process and oncostatic effect of melatonin were seen to be associated with extracellular-signal-regulated kinase (ERK) and stress-activated protein kinase/c-Jun NH (2)-terminal kinase (SAPK-JNK) signalling cascades in HeLa cells. In HT-29 and JURKAT cells, melatonin induced apoptosis via activation of p38 mitogen-activated protein kinases (p38), ERK and SAPK-JNK signalling pathways. In all three cell lines, the apoptotic event was triggered by the mammalian target of rapamycin (mTOR)-mediated activation of the downstream target rapamycininsensitive companion of mTOR (RICTOR) and/or regulatory-associated protein of mTOR (RAPTOR) proteins.

Conclusion: Our findings confirm that melatonin induces apoptosis through reactive oxygen speciesmediated dysregulated mitogen-activated protein kinase (MAPK) and mTOR signalling pathways in these cancer cell lines.

Keywords: Melatonin, apoptosis, cancer, reactive oxygen species, MAPK, mTOR.

Graphical Abstract

[1]
Global burden of disease cancer collaboration. The global burden of cancer 2013. JAMA Oncol 2015; 1:: 505-27.
[http://dx.doi.org/10.1001/jamaoncol.2015.0735]
[2]
GBD 2015 Risk Factors Collaborators. Global, regional, and national comparative risk assessment of 79 behavioural, environmental and occupational, and metabolic risks or clusters of risks, 1990-2015: A systematic analysis for the global burden of disease study 2015. Lancet 2016; 388(10053): 1659-724.
[http://dx.doi.org/10.1016/S0140-6736(16)31679-8] [PMID: 27733284]
[3]
Adams M, Jewell AP. The use of complementary and alternative medicine by cancer patients. Int Semin Surg Oncol 2007; 4: 10-7.
[http://dx.doi.org/10.1186/1477-7800-4-10] [PMID: 17470282]
[4]
Reiter RJ, Paredes SD, Manchester LC, Tan DX. Reducing oxidative/nitrosative stress: a newly-discovered genre for melatonin. Crit Rev Biochem Mol Biol 2009; 44(4): 175-200.
[http://dx.doi.org/10.1080/10409230903044914] [PMID: 19635037]
[5]
García-Santos G, Antolín I, Herrera F, et al. Melatonin induces apoptosis in human neuroblastoma cancer cells. J Pineal Res 2006; 41(2): 130-5.
[http://dx.doi.org/10.1111/j.1600-079X.2006.00342.x] [PMID: 16879318]
[6]
Zha L, Fan L, Sun G, et al. Melatonin sensitizes human hepatoma cells to endoplasmic reticulum stress-induced apoptosis. J Pineal Res 2012; 52(3): 322-31.
[http://dx.doi.org/10.1111/j.1600-079X.2011.00946.x] [PMID: 22225575]
[7]
Moreira AJ, Ordoñez R, Cerski CT, et al. Melatonin activates endoplasmic reticulum stress and apoptosis in rats with diethylnitrosamine-induced hepatocarcinogenesis. PLoS One 2015; 10(12)e0144517
[http://dx.doi.org/10.1371/journal.pone.0144517] [PMID: 26656265]
[8]
Liu H, Xu L, Wei JE, Xie MR, Wang SE, Zhou RX. Role of CD4+ CD25+ regulatory T cells in melatonin-mediated inhibition of murine gastric cancer cell growth in vivo and in vitro. Anat Rec (Hoboken) 2011; 294(5): 781-8.
[http://dx.doi.org/10.1002/ar.21361] [PMID: 21416626]
[9]
Kucharewicz K, Dudkowska M, Zawadzka A, et al. Simultaneous induction and blockade of autophagy by a single agent. Cell Death Dis 2018; 9(3): 353.
[http://dx.doi.org/10.1038/s41419-018-0383-6] [PMID: 29500364]
[10]
Sánchez DI, González-Fernández B, San-Miguel B, et al. Melatonin prevents deregulation of the sphingosine kinase/sphingosine 1-phosphate signaling pathway in a mouse model of diethylnitrosamine-induced hepatocellular carcinoma. J Pineal Res 2017; 62(1): e12369
[http://dx.doi.org/10.1111/jpi.12369] [PMID: 27696512]
[11]
Ferreira GM, Martinez M, Camargo ICC, Domeniconi RF, Martinez FE, Chuffa LGA. Melatonin attenuates Her-2, p38 MAPK, p-AKT, and mTOR levels in ovarian carcinoma of ethanol-preferring rats. J Cancer 2014; 5(9): 728-35.
[http://dx.doi.org/10.7150/jca.10196] [PMID: 25368672]
[12]
Girgert R, Bartsch C, Hill SM, Kreienberg R, Hanf V. Tracking the elusive antiestrogenic effect of melatonin: A new methodological approach. Neuroendocrinol Lett 2003; 24(6): 440-4.
[PMID: 15073572]
[13]
Martínez-Campa C, Alonso-González C, Mediavilla MD, et al. Melatonin inhibits both ER alpha activation and breast cancer cell proliferation induced by a metalloestrogen, cadmium. J Pineal Res 2006; 40(4): 291-6.
[http://dx.doi.org/10.1111/j.1600-079X.2006.00315.x] [PMID: 16635015]
[14]
Uguz AC, Cig B, Espino J, et al. Melatonin potentiates chemotherapy-induced cytotoxicity and apoptosis in rat pancreatic tumor cells. J Pineal Res 2012; 53(1): 91-8.
[http://dx.doi.org/10.1111/j.1600-079X.2012.00974.x] [PMID: 22288984]
[15]
Lissoni P, Rovelli F, Malugani F, Bucovec R, Conti A, Maestroni GJ. Anti-angiogenic activity of melatonin in advanced cancer patients. Neuroendocrinol Lett 2001; 22(1): 45-7.
[PMID: 11335879]
[16]
Alvarez-García V, González A, Alonso-González C, Martínez-Campa C, Cos S. Regulation of vascular endothelial growth factor by melatonin in human breast cancer cells. J Pineal Res 2013; 54(4): 373-80.
[http://dx.doi.org/10.1111/jpi.12007] [PMID: 23013414]
[17]
Borin TF, Arbab AS, Gelaleti GB, et al. Melatonin decreases breast cancer metastasis by modulating Rho-associated kinase protein-1 expression. J Pineal Res 2016; 60(1): 3-15.
[http://dx.doi.org/10.1111/jpi.12270] [PMID: 26292662]
[18]
Goncalves Ndo N, Rodrigues RV, Jardim-Perassi BV, et al. Molecular markers of angiogenesis and metastasis in lines of oral carcinoma after treatment with melatonin. Anticancer Agents Med Chem 2014; 14(9): 1302-11.
[http://dx.doi.org/10.2174/1871520614666140812110246] [PMID: 25323035]
[19]
Garcia-Mauriño S, Gonzalez-Haba MG, Calvo JR, et al. Melatonin enhances IL-2, IL-6, and IFN-gamma production by human circulating CD4+ cells: A possible nuclear receptor-mediated mechanism involving T helper type 1 lymphocytes and monocytes. J Immunol 1997; 159(2): 574-81.
[PMID: 9218571]
[20]
Hong Y, Won J, Lee Y, et al. Melatonin treatment induces interplay of apoptosis, autophagy, and senescence in human colorectal cancer cells. J Pineal Res 2014; 56(3): 264-74.
[http://dx.doi.org/10.1111/jpi.12119] [PMID: 24484372]
[21]
Bułdak RJ, Pilc-Gumuła K, Bułdak Ł, et al. Effects of ghrelin, leptin and melatonin on the levels of reactive oxygen species, antioxidant enzyme activity and viability of the HCT 116 human colorectal carcinoma cell line. Mol Med Rep 2015; 12(2): 2275-82.
[http://dx.doi.org/10.3892/mmr.2015.3599] [PMID: 25873273]
[22]
Batista AP, da Silva TG, Teixeira AA, et al. Ultrastructural aspects of melatonin cytotoxicity on Caco-2 cells in vitro. Micron 2014; 59: 17-23.
[http://dx.doi.org/10.1016/j.micron.2013.12.003] [PMID: 24530360]
[23]
León J, Casado J, Jiménez Ruiz SM, et al. Melatonin reduces endothelin-1 expression and secretion in colon cancer cells through the inactivation of FoxO-1 and NF-κβ. J Pineal Res 2014; 56(4): 415-26.
[http://dx.doi.org/10.1111/jpi.12131] [PMID: 24628039]
[24]
García-Navarro A, González-Puga C, Escames G, et al. Cellular mechanisms involved in the melatonin inhibition of HT-29 human colon cancer cell proliferation in culture. J Pineal Res 2007; 43(2): 195-205.
[http://dx.doi.org/10.1111/j.1600-079X.2007.00463.x] [PMID: 17645698]
[25]
Casado-Zapico S, Martín V, García-Santos G, et al. Regulation of the expression of death receptors and their ligands by melatonin in haematological cancer cell lines and in leukaemia cells from patients. J Pineal Res 2011; 50(3): 345-55.
[http://dx.doi.org/10.1111/j.1600-079X.2010.00850.x] [PMID: 21392090]
[26]
Margheri M, Pacini N, Tani A, et al. Combined effects of melatonin and all-trans retinoic acid and somatostatin on breast cancer cell proliferation and death: Molecular basis for the anticancer effect of these molecules. Eur J Pharmacol 2012; 681(1-3): 34-43.
[http://dx.doi.org/10.1016/j.ejphar.2012.02.011] [PMID: 22532966]
[27]
Jain P, Silva A, Han HJ, et al. Overcoming resistance to single-agent therapy for oncogenic BRAF gene fusions via combinatorial targeting of MAPK and PI3K/mTOR signaling pathways. Oncotarget 2017; 8(49): 84697-713.
[http://dx.doi.org/10.18632/oncotarget.20949] [PMID: 29156677]
[28]
Wang JH1. Zhou YJ, He P, Chen BY.. Role of mitogen-activated protein kinase pathways during Escherichia coli-induced apoptosis in U937 cells. Apoptosis 2007; 12: 375-85.
[http://dx.doi.org/10.1007/s10495-006-0623-6] [PMID: 17191113]
[29]
Adhikari S, Bhatia M. H2S-induced pancreatic acinar cell apoptosis is mediated via JNK and p38 MAP kinase. J Cell Mol Med 2008; 12(4): 1374-83.
[http://dx.doi.org/10.1111/j.1582-4934.2008.00318.x] [PMID: 18373739]
[30]
Li W, Fan M, Chen Y, et al. Melatonin induces cell apoptosis in AGS cells through the activation of JNK and P38 MAPK and the suppression of nuclear factor-kappa B: A novel therapeutic implication for gastric cancer. Cell Physiol Biochem 2015; 37(6): 2323-38.
[http://dx.doi.org/10.1159/000438587] [PMID: 26645893]
[31]
Zhou Q, Gui S, Zhou Q, Wang Y. Melatonin inhibits the migration of human lung adenocarcinoma A549 cell lines involving JNK/MAPK pathway. PLoS One 2014; 9(7): e101132
[http://dx.doi.org/10.1371/journal.pone.0101132] [PMID: 24992189]
[32]
Penna I, Molla A, Grazia G, et al. Primary cross-resistance to BRAFV600E-, MEK1/2- and PI3K/mTOR-specific inhibitors in BRAF-mutant melanoma cells counteracted by dual pathway blockade. Oncotarget 2016; 7(4): 3947-65.
[http://dx.doi.org/10.18632/oncotarget.6600] [PMID: 26678033]
[33]
Lamhamedi-Cherradi SE, Menegaz BA, Ramamoorthy V, et al. IGF-1R and mTOR Blockade: Novel Resistance mechanisms and synergistic drug combinations for Ewing sarcoma. J Natl Cancer Inst 2016; 108(12): 1-12.
[http://dx.doi.org/10.1093/jnci/djw182] [PMID: 27576731]
[34]
Mondesire WH, Jian W, Zhang H, et al. Targeting mammalian target of rapamycin synergistically enhances chemotherapy-induced cytotoxicity in breast cancer cells. Clin Cancer Res 2004; 10(20): 7031-42.
[http://dx.doi.org/10.1158/1078-0432.CCR-04-0361] [PMID: 15501983]
[35]
Zhao L, Teng B, Wen L, et al. mTOR inhibitor AZD8055 inhibits proliferation and induces apoptosis in laryngeal carcinoma. Int J Clin Exp Med 2014; 7(2): 337-47.
[PMID: 24600487]
[36]
Wenzel U, Nickel A, Daniel H. Melatonin potentiates flavone-induced apoptosis in human colon cancer cells by increasing the level of glycolytic end products. Int J Cancer 2005; 116(2): 236-42.
[http://dx.doi.org/10.1002/ijc.20837] [PMID: 15800915]
[37]
Sánchez-Hidalgo M, Lee M, de la Lastra CA, Guerrero JM, Packham G. Melatonin inhibits cell proliferation and induces caspase activation and apoptosis in human malignant lymphoid cell lines. J Pineal Res 2012; 53(4): 366-73.
[http://dx.doi.org/10.1111/j.1600-079X.2012.01006.x] [PMID: 22582944]
[38]
Papazisis KT, Kouretas D, Geromichalos GD, et al. Effects of melatonin on proliferation of cancer cell lines. J Pineal Res 1998; 25(4): 211-8.
[http://dx.doi.org/10.1111/j.1600-079X.1998.tb00390.x] [PMID: 9885990]
[39]
Farriol M, Venereo Y, Orta X, Castellanos JM, Segovia-Silvestre T. In vitro effects of melatonin on cell proliferation in a colon adenocarcinoma line. J Appl Toxicol 2000; 20(1): 21-4.
[http://dx.doi.org/10.1002/(SICI)1099-1263(200001/02)20:1<21::AID-JAT623>3.0.CO;2-M] [PMID: 10641013]
[40]
He W, Li X, Xia S. Lupeol triterpene exhibits potent antitumor effects in A427 human lung carcinoma cells via mitochondrial mediated apoptosis, ROS generation, loss of mitochondrial membrane potential and downregulation of m-TOR/PI3Ksol; AKT signalling pathway. J BUON 2018; 23(3): 635-40.
[PMID: 30003730]
[41]
Xia S, Miao Y, Liu S. Withaferin A induces apoptosis by ROS-dependent mitochondrial dysfunction in human colorectal cancer cells. Biochem Biophys Res Commun 2018; 503(4): 2363-9.
[http://dx.doi.org/10.1016/j.bbrc.2018.06.162] [PMID: 29966656]
[42]
Childs AC, Phaneuf SL, Dirks AJ, Phillips T, Leeuwenburgh C. Doxorubicin treatment in vivo causes cytochrome C release and cardiomyocyte apoptosis, as well as increased mitochondrial efficiency, superoxide dismutase activity, and Bcl-2: Bax ratio. Cancer Res 2002; 62(16): 4592-8.
[PMID: 12183413]
[43]
Cabrera J, Negrín G, Estévez F, Loro J, Reiter RJ, Quintana J. Melatonin decreases cell proliferation and induces melanogenesis in human melanoma SK-MEL-1 cells. J Pineal Res 2010; 49(1): 45-54.
[http://dx.doi.org/10.1111/j.1600-079X.2010.00765.x] [PMID: 20459460]
[44]
Hill SM, Blask DE. Effects of the pineal hormone melatonin on the proliferation and morphological characteristics of human breast cancer cells (MCF-7) in culture. Cancer Res 1988; 48(21): 6121-6.
[PMID: 3167858]
[45]
Petranka J, Baldwin W, Biermann J, Jayadev S, Barrett JC, Murphy E. The oncostatic action of melatonin in an ovarian carcinoma cell line. J Pineal Res 1999; 26(3): 129-36.
[http://dx.doi.org/10.1111/j.1600-079X.1999.tb00574.x] [PMID: 10231725]
[46]
Yun CW, Kim S, Lee JH, Lee SH. Melatonin promotes apoptosis of colorectal cancer cells via superoxide-mediated ER stress by inhibiting cellular prion protein expression. Anticancer Res 2018; 38: 3951-60.
[47]
Katagiri K, Matsuzawa A, Ichijo H. Regulation of apoptosis signal-regulating kinase 1 in redox signaling. Methods Enzymol 2010; 474: 277-88.
[48]
Friday BB, Adjei AA. Advances in targeting the Ras/Raf/MEK/Erk mitogen-activated protein kinase cascade with MEK inhibitors for cancer therapy. Clin Cancer Res 2008; 14(2): 342-6.
[http://dx.doi.org/10.1158/1078-0432.CCR-07-4790] [PMID: 18223206]
[49]
Oh HN, Seo JH, Lee MH, et al. Oridonin induces apoptosis in oral squamous cell carcinoma probably through the generation of reactive oxygen species and the p38/JNK MAPK pathway. Int J Oncol 2018; 4319.
[http://dx.doi.org/10.3892/ijo.2018.4319] [PMID: 29568920]
[50]
Cho HJ, Ahn KC, Choi JY, et al. Luteolin acts as a radiosensitizer in non-small cell lung cancer cells by enhancing apoptotic cell death through activation of a p38/ROS/caspase cascade. Int J Oncol 2015; 46(3): 1149-58.
[http://dx.doi.org/10.3892/ijo.2015.2831] [PMID: 25586525]
[51]
Lieberthal W, Levine JS. The role of the mammalian target of rapamycin (mTOR) in renal disease. J Am Soc Nephrol 2009; 20(12): 2493-502.
[http://dx.doi.org/10.1681/ASN.2008111186] [PMID: 19875810]
[52]
Kim DH, Sarbassov DD, Ali SM, et al. mTOR interacts with raptor to form a nutrient-sensitive complex that signals to the cell growth machinery. Cell 2002; 110(2): 163-75.
[http://dx.doi.org/10.1016/S0092-8674(02)00808-5] [PMID: 12150925]
[53]
Blagosklonny MV. Aging: ROS or TOR. Cell Cycle 2008; 7(21): 3344-54.
[http://dx.doi.org/10.4161/cc.7.21.6965] [PMID: 18971624]
[54]
Chen D, Lin X, Zhang C, et al. Dual PI3K/mTOR inhibitor BEZ235 as a promising therapeutic strategy against paclitaxel-resistant gastric cancer via targeting PI3K/Akt/mTOR pathway. Cell Death Dis 2018; 9(2): 123-34.
[http://dx.doi.org/10.1038/s41419-017-0132-2] [PMID: 29374144]
[55]
Ghosh R, Gilda JE, Gomes AV. The necessity of and strategies for improving confidence in the accuracy of western blots. Expert Rev Proteomics 2014; 11(5): 549-60.
[http://dx.doi.org/10.1586/14789450.2014.939635] [PMID: 25059473]

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