Activation of matrix metalloproteinase-9 (MMP-9) by neurotensin promotes cell invasion and migration through ERK pathway in gastric cancer
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Neurotensin (NT) is distributed throughout the brain and gastrointestinal tract. Although the relationship between NT and matrix metalloproteinase-9 (MMP-9) activity in gastric cancer has not been reported, the elevation of MMP-9 and NT is reported in the breast, lung, prostate, and gastric cancer. The aim of our study is to investigate the relationship between NT and MMP-9 activity and the underlying signaling mechanism in gastric cancer cell lines. Commercial ELISA kits were used for estimation of NT and MMP-9 expression, and fluorescence resonance energy transfer (FRET) assay was used for measurement of MMP-9 activity. Cell migration and invasion were determined by wound healing and transwell assay. The expression of signaling proteins was measured by Western blotting. Our study reveals a positive correlation between increased plasma NT and MMP-9 activity in both of patient’s serum and gastric cancer cell lines. A dose-dependent elevation of MMP-9 activity was observed by NT treatment in gastric cancer cells (MKN-1 and MKN-45) compared to untreated gastric cancer and normal epithelial cell (HFE-145). Moreover, NT-mediated migration and invasion were observed in gastric cancer cells unlike in normal cell. The signaling mechanism of NT in gastric cancer cells was confirmed in protein kinase C (PKC), extracellular-signal regulated kinase (ERK), and phosphatidylinositol 3-kinase (PI3K) pathway. In addition, pretreatment of gastric cancer cells with NTR1 inhibitor SR48692 was shown to significantly inhibit the NT-mediated MMP-9 activity, cell invasion, and migration. Our finding illustrated NTR1 could be a possible therapeutic target for gastric cancer.
KeywordsNeurotensin NTR1 ELISA MMP-9 Invasion ERK
This study was supported by the Creative Fusion Research Program through the Creative Allied Project funded by Korea Research Council of Science and Technology (CAP-12-1-KIST) and Korea Institute of Science and Technology (KIST) Institutional Program (project no. 2E25360).
Conflicts of interest
- 10.Reubi JC, Waser B, Schmassmann A, Laissue JA. Receptor autoradiographic evaluation of cholecystokinin, neurotensin, somatostatin and vasoactive intestinal peptide receptors in gastro-intestinal adenocarcinoma samples: where are they really located? Int J Cancer. 1999;81:376–86.CrossRefPubMedGoogle Scholar
- 11.Shimizu S, Tsukada J, Sugimoto T, Kikkawa N, Sasaki K, Chazono H, et al. Identification of a novel therapeutic target for head and neck squamous cell carcinomas: a role for the neurotensin-neurotensin receptor 1 oncogenic signaling pathway. Int J Cancer. 2008;123:1816–23.CrossRefPubMedGoogle Scholar
- 25.Maatta M, Soini Y, Liakka A, Autio-Harmainen H. Differential expression of matrix metalloproteinase (MMP)-2, MMP-9, and membrane type 1-MMP in hepatocellular and pancreatic adenocarcinoma: implications for tumor progression and clinical prognosis. Clin Cancer Res. 2000;6:2726–34.PubMedGoogle Scholar
- 29.Liao A, Wang W, Sun D, Jiang Y, Tian S, Li J, Yang X, Shi R. Bone morphogenetic protein 2 mediates epithelial-mesenchymal transition via AKT and ERK signaling pathways in gastric cancer. Tumour Biology: the Journal of the International Society for Oncodevelopmental Biology and Medicine. 2014.Google Scholar
- 33.FernandezFernandez L, Tejero E, Tieso A, Rabadan L, Munoz M, Santos I. Receiver operating characteristic (ROC) curve analysis of the tumour markers CEA, CA 19-9 and CA 72-4 in gastric cancer. Int Surg. 1996;81:400–2.Google Scholar
- 37.Lee CW, Lin CC, Lin WN, Liang KC, Luo SF, Wu CB, et al. TNF-alpha induces MMP-9 expression via activation of SRC/EGFR, PDGFR/PI3K/Akt cascade and promotion of NF-kappab/p300 binding in human tracheal smooth muscle cells. Am J Physiol Lung Cell Mol Physiol. 2007;292:L799–812.CrossRefPubMedGoogle Scholar
- 41.di Florio A, Sancho V, Moreno P, Delle Fave G, Jensen RT. Gastrointestinal hormones stimulate growth of foregut neuroendocrine tumors by transactivating the EGF receptor. BBA Mol Cell Res. 2013;1833:573–82.Google Scholar