Inhibition of matrix metalloproteinase-9 and nuclear factor kappa B contribute to melatonin prevention of motility and invasiveness in HepG2 liver cancer cells
Raquel Ordoñez
Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), León, Spain
Institute of Biomedicine (IBIOMED), University of León, León, Spain
Search for more papers by this authorSara Carbajo-Pescador
Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), León, Spain
Institute of Biomedicine (IBIOMED), University of León, León, Spain
Search for more papers by this authorNéstor Prieto-Dominguez
Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), León, Spain
Institute of Biomedicine (IBIOMED), University of León, León, Spain
Search for more papers by this authorAndrés García-Palomo
Institute of Biomedicine (IBIOMED), University of León, León, Spain
Service of Oncology, Hospital of León, Complejo Asistencial Universitario de León, León, Spain
Search for more papers by this authorCorresponding Author
Javier González-Gallego
Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), León, Spain
Institute of Biomedicine (IBIOMED), University of León, León, Spain
Address reprint requests to Javier González-Gallego, Institute of Biomedicine, University of León, 24071-León, Spain.
E-mail: [email protected]
Search for more papers by this authorJosé L. Mauriz
Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), León, Spain
Institute of Biomedicine (IBIOMED), University of León, León, Spain
Search for more papers by this authorRaquel Ordoñez
Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), León, Spain
Institute of Biomedicine (IBIOMED), University of León, León, Spain
Search for more papers by this authorSara Carbajo-Pescador
Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), León, Spain
Institute of Biomedicine (IBIOMED), University of León, León, Spain
Search for more papers by this authorNéstor Prieto-Dominguez
Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), León, Spain
Institute of Biomedicine (IBIOMED), University of León, León, Spain
Search for more papers by this authorAndrés García-Palomo
Institute of Biomedicine (IBIOMED), University of León, León, Spain
Service of Oncology, Hospital of León, Complejo Asistencial Universitario de León, León, Spain
Search for more papers by this authorCorresponding Author
Javier González-Gallego
Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), León, Spain
Institute of Biomedicine (IBIOMED), University of León, León, Spain
Address reprint requests to Javier González-Gallego, Institute of Biomedicine, University of León, 24071-León, Spain.
E-mail: [email protected]
Search for more papers by this authorJosé L. Mauriz
Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), León, Spain
Institute of Biomedicine (IBIOMED), University of León, León, Spain
Search for more papers by this authorAbstract
Hepatocellular carcinoma (HCC) is one of the most lethal human cancers worldwide because of its high incidence and its metastatic potential. Extracellular matrix degradation by matrix metalloproteinases (MMPs) has been connected with cancer cell invasion, and it has been suggested that inhibition of MMPs by synthetic and natural inhibitors may be of great importance in the HCC therapies. Melatonin, the main product of the pineal gland, exerts antiproliferative, proapoptotic, and antiangiogenic properties in HepG2 human hepatocellular cells, and exhibits anti-invasive and antimetastatic activities by suppressing the enzymatic activity of MMP-9 in different tumor types. However, the underlying mechanism of anti-invasive activity in HCC models has not been fully elucidated. Here, we demonstrate that 1 mm melatonin dosage reduced in IL-1β-induced HepG2 cells MMP-9 gelatinase activity and inhibited cell invasion and motility through downregulation of MMP-9 gene expression and upregulation of the MMP-9-specific inhibitor tissue inhibitor of metalloproteinases (TIMP)-1. No significant changes were observed in the expression and activity of MMP-2, the other proteinase implicated in matrix collagen degradation, and its tissue inhibitor, TIMP-2. Also, melatonin significantly suppressed IL-1β-induced nuclear factor-kappaB (NF-κB) translocation and transcriptional activity. In summary, we demonstrate that melatonin modulates motility and invasiveness of HepG2 cell in vitro through a molecular mechanism that involves TIMP-1 upregulation and attenuation of MMP-9 expression and activity via NF-κB signal pathway inhibition.
References
- 1Jemal A, Bray F, Center MM et al. Global cancer statistics. CA Cancer J Clin 2011; 612: 69–90.
10.3322/caac.20107 Google Scholar
- 2El-Serag HB. Hepatocellular carcinoma. N Engl J Med 2011; 365: 1118–1127.
- 3Marsh JW, Dvorchik I, Subotin M et al. The prediction of risk of recurrence and time to recurrence of hepatocellular carcinoma after orthotopic liver transplantation: a pilot study. Hepatology 1997; 26: 444–450.
- 4Okada H, Kamino Y, Shimo M et al. Metastatic hepatocellular carcinoma of the maxillary sinus: a rare autopsy case without lung metastasis and a review. Int J Oral Maxillofac Surg 2003; 32: 97–100.
- 5Thuluvath PJ. Vascular invasion is the most important predictor of survival in HCC, but how do we find it? J Clin Gastroenterol 2009; 43: 101–102.
- 6Shen B, Chu ES, Zhao G et al. PPARgamma inhibits hepatocellular carcinoma metastases in vitro and in mice. Br J Cancer 2012; 106: 1486–1494.
- 7Fidler IJ. The pathogenesis of cancer metastasis: the ‘seed and soil’ hypothesis revisited. Nat Rev Cancer 2003; 3: 453–458.
- 8Westermarck J, Kahari VM. Regulation of matrix metalloproteinase expression in tumor invasion. FASEB J 1999; 13: 781–792.
- 9Kader AK, Liu J, Shao L et al. Matrix metalloproteinase polymorphisms are associated with bladder cancer invasiveness. Clin Cancer Res 2007; 13: 2614–2620.
- 10Arii S, Mise M, Harada T et al. Overexpression of matrix metalloproteinase 9 gene in hepatocellular carcinoma with invasive potential. Hepatology 1996; 24: 316–322.
- 11Yu H, Pan C, Zhao S et al. Resveratrol inhibits tumor necrosis factor-alpha-mediated matrix metalloproteinase-9 expression and invasion of human hepatocellular carcinoma cells. Biomed Pharmacother 2008; 62: 366–372.
- 12Stamenkovic I. Matrix metalloproteinases in tumor invasion and metastasis. Semin Cancer Biol 2000; 10: 415–433.
- 13Bode W. Structural basis of matrix metalloproteinase function. Biochem Soc Symp 2003; 70: 1–14.
- 14Bourboulia D, Stetler-Stevenson WG. Matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs): positive and negative regulators in tumor cell adhesion. Semin Cancer Biol 2010; 20: 161–168.
- 15Nasatzky E, Rubinstein Y, Goultschin J et al. The role of Matrix Metaloproteinases in the progression of periodontitis, and the use of specific inhibitors to these enzymes in the treatment of the periodontal disease. Refuat Hapeh Vehashinayim 2003; 20: 81.
- 16Kim YH, Kwon HJ, Kim DS. Matrix metalloproteinase 9 (MMP-9)-dependent processing of betaig-h3 protein regulates cell migration, invasion, and adhesion. J Biol Chem 2012; 287: 38957–38969.
- 17Yang YN, Wang F, Zhou W et al. TNF-alpha stimulates MMP-2 and MMP-9 activities in human corneal epithelial cells via the activation of FAK/ERK signaling. Ophthalmic Res 2012; 48: 165–170.
- 18Arsura M, Cavin LG. Nuclear factor-kappaB and liver carcinogenesis. Cancer Lett 2005; 229: 157–169.
- 19Maeda S, Omata M. Inflammation and cancer: role of nuclear factor-kappaB activation. Cancer Sci 2008; 99: 836–842.
- 20Cuevas MJ, Almar M, García-González JC et al. Changes in oxidative stress markers and NF-kappa B activation induced by sprint exercise. Free Radical Res 2005; 39: 431–439.
- 21Veneroso C, Tuñon MJ, González-Gallego J et al. Melatonin reduces cardiac inflammatory injury induced by acute exercise. J Pineal Res 2009; 47: 184–191.
- 22Lawrence T. The nuclear factor NF-kappaB pathway in inflammation. Cold Spring Harb Perspect Biol 2009; 1: a001651.
- 23Pan M, Song YL, Xu JM et al. Melatonin ameliorates nonalcoholic fatty liver induced by high-fat diet in rats. J Pineal Res 2006; 41: 79–84.
- 24Subramanian P, Mirunalini S, Pandi-Perumal SR et al. Melatonin treatment improves the antioxidant status and decreases lipid content in brain and liver of rats. Eur J Pharmacol 2007; 571: 116–119.
- 25Tahan V, Atug O, Akin H et al. Melatonin ameliorates methionine- and choline-deficient diet-induced nonalcoholic steatohepatitis in rats. J Pineal Res 2009; 46: 401–407.
- 26Thong-Ngam D, Samuhasaneeto S, Kulaputana O et al. N-acetylcysteine attenuates oxidative stress and liver pathology in rats with non-alcoholic steatohepatitis. World J Gastroenterol 2007; 13: 5127–5132.
- 27Cabrera J, Negrin G, Estevez F et al. Melatonin decreases cell proliferation and induces melanogenesis in human melanoma SK-MEL-1 cells. J Pineal Res 2010; 49: 45–54.
- 28Chiu CC, Chen JY, Lin KL et al. p38 MAPK and NF-kappaB pathways are involved in naphtho[1,2-b] furan-4,5-dione induced anti-proliferation and apoptosis of human hepatoma cells. Cancer Lett 2010; 295: 92–99.
- 29Cini G, Neri B, Pacini A et al. Antiproliferative activity of melatonin by transcriptional inhibition of cyclin D1 expression: a molecular basis for melatonin-induced oncostatic effects. J Pineal Res 2005; 39: 12–20.
- 30Farriol M, Venereo Y, Orta X et al. In vitro effects of melatonin on cell proliferation in a colon adenocarcinoma line. J Appl Toxicol 2000; 20: 21–24.
10.1002/(SICI)1099-1263(200001/02)20:1<21::AID-JAT623>3.0.CO;2-M CASPubMedWeb of Science®Google Scholar
- 31Futagami M, Sato S, Sakamoto T et al. Effects of melatonin on the proliferation and cis-diamminedichloroplatinum (CDDP) sensitivity of cultured human ovarian cancer cells. Gynecol Oncol 2001; 82: 544–549.
- 32Garcia-Navarro A, Gonzalez-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: 195–205.
- 33Garcia-Santos G, Antolin I, Herrera F et al. Melatonin induces apoptosis in human neuroblastoma cancer cells. J Pineal Res 2006; 41: 130–135.
- 34Gonzalez A, Del Castillo-Vaquero A, Miro-Moran A et al. Melatonin reduces pancreatic tumor cell viability by altering mitochondrial physiology. J Pineal Res 2010; 50: 250–260.
- 35Hill 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: 6121–6126.
- 36Lissoni P, Rovelli F, Malugani F et al. Anti-angiogenic activity of melatonin in advanced cancer patients. Neuro Endocrinol Lett 2001; 22: 45–47.
- 37Cui P, Yu M, Peng X et al. Melatonin prevents human pancreatic carcinoma cell PANC-1-induced human umbilical vein endothelial cell proliferation and migration by inhibiting vascular endothelial growth factor expression. J Pineal Res 2012; 52: 236–243.
- 38Kim KJ, Choi JS, Kang I et al. Melatonin suppresses tumor progression by reducing angiogenesis stimulated by HIF-1 in a mouse tumor model. J Pineal Res 2012; 54: 264–270.
- 39Swarnakar S, Paul S, Singh LP et al. Matrix metalloproteinases in health and disease: regulation by melatonin. J Pineal Res 2011; 50: 8–20.
- 40Wang J, Hao H, Yao L et al. Melatonin suppresses migration and invasion via inhibition of oxidative stress pathway in glioma cells. J Pineal Res 2012; 53: 180–187.
- 41Zhang HJ, Yao DF, Yao M et al. Annexin A2 silencing inhibits invasion, migration, and tumorigenic potential of hepatoma cells. World J Gastroenterol 2013; 19: 3792–3801.
- 42Ortiz-Lopez L, Morales-Mulia S, Ramirez-Rodriguez G et al. ROCK-regulated cytoskeletal dynamics participate in the inhibitory effect of melatonin on cancer cell migration. J Pineal Res 2009; 46: 15–21.
- 43Cos S, Fernandez R, Guezmes A et al. Influence of melatonin on invasive and metastatic properties of MCF-7 human breast cancer cells. Cancer Res 1998; 58: 4383–4390.
- 44Carbajo-Pescador S, Ordonez R, Benet M et al. Inhibition of VEGF expression through blockade of Hif1alpha and STAT3 signalling mediates the anti-angiogenic effect of melatonin in HepG2 liver cancer cells. Br J Cancer 2013; 109: 83–91.
- 45Carbajo-Pescador S, Steinmetz C, Kashyap A et al. Melatonin induces transcriptional regulation of Bim by FoxO3a in HepG2 cells. Br J Cancer 2013; 108: 442–449.
- 46Carbajo-Pescador S, Garcia-Palomo A, Martin-Renedo J et al. Melatonin modulation of intracellular signaling pathways in hepatocarcinoma HepG2 cell line: role of the MT1 receptor. J Pineal Res 2011; 51: 463–471.
- 47Carbajo-Pescador S, Martin-Renedo J, Garcia-Palomo A et al. Changes in the expression of melatonin receptors induced by melatonin treatment in hepatocarcinoma HepG2 cells. J Pineal Res 2009; 47: 330–338.
- 48Martin-Renedo J, Mauriz JL, Jorquera F et al. Melatonin induces cell cycle arrest and apoptosis in hepatocarcinoma HepG2 cell line. J Pineal Res 2008; 45: 532–540.
- 49Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 2001; 25: 402–408.
- 50Apte RN, Dotan S, Elkabets M et al. The involvement of IL-1 in tumorigenesis, tumor invasiveness, metastasis and tumor-host interactions. Cancer Metastasis Rev 2006; 25: 387–408.
- 51Katyal S, Oliver JH3rd, Peterson MS et al. Extrahepatic metastases of hepatocellular carcinoma. Radiology 2000; 216: 698–703.
- 52Noel A, Gutierrez-Fernandez A, Sounni NE et al. New and paradoxical roles of matrix metalloproteinases in the tumor microenvironment. Front Pharmacol 2012; 3: 140.
- 53Coronato S, Laguens G, di Girolamo V. Role of metalloproteinases and their inhibitors in tumors. Medicina (B Aires) 2012; 72: 495–502.
- 54Hadler-Olsen E, Winberg JO, Uhlin-Hansen L. Matrix metalloproteinases in cancer: their value as diagnostic and prognostic markers and therapeutic targets. Tumour Biol 2013; 34: 2041–2051.
- 55Hahm JH, Kim S, Paik YK. Endogenous cGMP regulates adult longevity via the insulin signaling pathway in Caenorhabditis elegans. Aging Cell 2009; 8: 473–483.
- 56Seely D, Wu P, Fritz H et al. Melatonin as adjuvant cancer care with and without chemotherapy: a systematic review and meta-analysis of randomized trials. Integr Cancer Ther 2012; 11: 293–303.
- 57Wang YM, Jin BZ, Ai F et al. The efficacy and safety of melatonin in concurrent chemotherapy or radiotherapy for solid tumors: a meta-analysis of randomized controlled trials. Cancer Chemother Pharmacol 2012; 69: 1213–1220.
- 58Papazisis KT, Kouretas D, Geromichalos GD et al. Effects of melatonin on proliferation of cancer cell lines. J Pineal Res 1998; 25: 211–218.
- 59Petranka J, Baldwin W, Biermann J et al. The oncostatic action of melatonin in an ovarian carcinoma cell line. J Pineal Res 1999; 26: 129–136.
- 60Shiu SY, Li L, Xu JN et al. Melatonin-induced inhibition of proliferation and G1/S cell cycle transition delay of human choriocarcinoma JAr cells: possible involvement of MT2 (MEL1B) receptor. J Pineal Res 1999; 27: 183–192.
- 61Cutando A, Aneiros-Fernandez J, Aneiros-Cachaza J et al. Melatonin and cancer: current knowledge and its application to oral cavity tumours. J Oral Pathol Med 2011; 40: 593–597.
- 62Nee LE, McMorrow T, Campbell E et al. TNF-alpha and IL-1beta-mediated regulation of MMP-9 and TIMP-1 in renal proximal tubular cells. Kidney Int 2004; 66: 1376–1386.
- 63Paul S, Sharma AV, Mahapatra PD et al. Role of melatonin in regulating matrix metalloproteinase-9 via tissue inhibitors of metalloproteinase-1 during protection against endometriosis. J Pineal Res 2008; 44: 439–449.
- 64Tai SH, Chen HY, Lee EJ et al. Melatonin inhibits postischemic matrix metalloproteinase-9 (MMP-9) activation via dual modulation of plasminogen/plasmin system and endogenous MMP inhibitor in mice subjected to transient focal cerebral ischemia. J Pineal Res 2010; 49: 332–341.
- 65Mao L, Yuan L, Slakey LM et al. Inhibition of breast cancer cell invasion by melatonin is mediated through regulation of the p38 mitogen-activated protein kinase signaling pathway. Breast Cancer Res 2010; 12: R107.
- 66Qin W, Lu W, Li H et al. Melatonin inhibits IL1beta-induced MMP9 expression and activity in human umbilical vein endothelial cells by suppressing NF-kappaB activation. J Endocrinol 2012; 214: 145–153.
- 67Rudra DS, Pal U, Maiti NC et al. Melatonin inhibits matrix metalloproteinase-9 activity by binding to its active site. J Pineal Res 2013; 54: 398–405.
- 68Ogawa K, Chen F, Kuang C et al. Suppression of matrix metalloproteinase-9 transcription by transforming growth factor-beta is mediated by a nuclear factor-kappaB site. Biochem J 2004; 381: 413–422.
- 69Tobar N, Villar V, Santibanez JF. ROS-NFkappaB mediates TGF-beta1-induced expression of urokinase-type plasminogen activator, matrix metalloproteinase-9 and cell invasion. Mol Cell Biochem 2010; 340: 195–202.
- 70Liu P, Kimmoun E, Legrand A et al. Activation of NF-kappa B, AP-1 and STAT transcription factors is a frequent and early event in human hepatocellular carcinomas. J Hepatol 2002; 37: 63–71.
- 71Chang CC, Tien CH, Lee EJ et al. Melatonin inhibits matrix metalloproteinase-9 (MMP-9) activation in the lipopolysaccharide (LPS)-stimulated RAW 264.7 and BV2 cells and a mouse model of meningitis. J Pineal Res 2012; 53: 188–197.
- 72Yang YI, Lee KT, Park HJ et al. Tectorigenin sensitizes paclitaxel-resistant human ovarian cancer cells through downregulation of the Akt and NFkappaB pathway. Carcinogenesis 2012; 33: 2488–2498.
- 73Kar S, Palit S, Ball WB et al. Carnosic acid modulates Akt/IKK/NF-kappaB signaling by PP2A and induces intrinsic and extrinsic pathway mediated apoptosis in human prostate carcinoma PC-3 cells. Apoptosis 2012; 17: 735–747.
- 74Lin KL, Tsai PC, Hsieh CY et al. Antimetastatic effect and mechanism of ovatodiolide in MDA-MB-231 human breast cancer cells. Chem Biol Interact 2011; 194: 148–158.
- 75Chiang IT, Liu YC, Wang WH et al. Sorafenib inhibits TPA-induced MMP-9 and VEGF expression via suppression of ERK/NF-kappaB pathway in hepatocellular carcinoma cells. In Vivo 2012; 26: 671–681.
- 76Shishodia S, Majumdar S, Banerjee S et al. Ursolic acid inhibits nuclear factor-kappaB activation induced by carcinogenic agents through suppression of IkappaBalpha kinase and p65 phosphorylation: correlation with down-regulation of cyclooxygenase 2, matrix metalloproteinase 9, and cyclin D1. Cancer Res 2003; 63: 4375–4383.
- 77Sethi G, Ahn KS, Pandey MK et al. Celastrol, a novel triterpene, potentiates TNF-induced apoptosis and suppresses invasion of tumor cells by inhibiting NF-kappaB-regulated gene products and TAK1-mediated NF-kappaB activation. Blood 2007; 109: 2727–2735.
- 78Buhrmann C, Mobasheri A, Busch F et al. Curcumin modulates nuclear factor kappaB (NF-kappaB)-mediated inflammation in human tenocytes in vitro: role of the phosphatidylinositol 3-kinase/Akt pathway. J Biol Chem 2011; 286: 28556–28566.
- 79Aggarwal BB, Shishodia S. Molecular targets of dietary agents for prevention and therapy of cancer. Biochem Pharmacol 2006; 71: 1397–1421.
- 80Liang JA, Wu SL, Lo HY et al. Vanillin inhibits matrix metalloproteinase-9 expression through down-regulation of nuclear factor-kappaB signaling pathway in human hepatocellular carcinoma cells. Mol Pharmacol 2009; 75: 151–157.
- 81Huang CS, Fan YE, Lin CY et al. Lycopene inhibits matrix metalloproteinase-9 expression and down-regulates the binding activity of nuclear factor-kappa B and stimulatory protein-1. J Nutr Biochem 2007; 18: 449–456.