Abstract
The aim of the study was to investigate the interaction between esophageal carcinoma cells with different differentiation degree and esophageal carcinoma-related lymphatic endothelial cells. Different lymphatic endothelial cell conditioned mediums were used to cultivate well-differentiated esophageal carcinoma EC9706 cells and poorly differentiated esophageal carcinoma KYSE150 cells, and immunocytochemistry and Western blot analyses were applied to detect the expression of MMP-9 protein and TIMP-2 protein in each group; in situ hybridization and RT-PCR methods were used to detect the expression of MMP-9 and TIMP-2 mRNA in each group; CCK-8 method was used to detect cell proliferation in each group; and transwell method was utilized to detect cell invasiveness in each group. Through constructing the transplanted tumor model of esophageal carcinoma of nude mice, the D2-40 and LYVE-1 immunohistochemical staining was performed on transplanted tumors and surrounding tissues, lymphatic microvessels were marked, and lymphatic microvessel density (LMVD) was measured. The expression of MMP-9 protein and mRNA in experimental group was significantly higher than that in control groups (P < 0.05); TIMP-2 protein and mRNA expression in experimental group was significantly lower than that in control groups (P < 0.05); cell proliferation ability and invasiveness ability in experimental group were significantly higher than those in control groups (P < 0.05); LMVD-marked D2-40 and LMVD-marked LYVE-1 of transplanted tumor tissue in the experimental group were significantly higher than those in control groups (P < 0.05). The esophageal squamous carcinoma-related lymphatic microvessel could promote the proliferation and invasive ability of esophageal squamous carcinoma cells in vitro. It had different effects on esophageal carcinoma cells with different differentiation degree and had more obvious influence on poorly differentiated esophageal carcinoma cells, which may be related to the up-regulated MMP-2 expression and down-regulated TIMP-2 expression of esophageal carcinoma cells. The esophageal squamous carcinoma-related lymphatic microvessel endothelial cells could promote the growth of esophageal carcinoma-transplanted tumor of nude mice and lymphangiogenesis.
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References
Song J, Su H, Zhou YY, et al. Prognostic value of matrix metalloproteinase 9 expression in breast cancer patients: a meta-analysis. Asian Pac J Cancer Prev. 2013;14(3):1615–21.
Zhu M, Zhang N, He S. MicroRNA-106a targets TIMP2 to regulate invasion and metastasis of gastric cancer. FEBS Lett. 2013;14(57):93–6.
Zebrowska A, Narbutt J, Sysa-Jedrzejowska A, et al. The imbalance between metalloproteinases and their tissue inhibitors is involved in the pathogenesis of dermatitis herpetiformis. Mediat Inflamm. 2005;25(6):373–9.
Mellick AS, Blackmore D, Weinstein SR. An assessment of MMP and TIMP gene expression in cell lines and stroma—tumour differences in microdissected breast cancer biopsies. Tumour Biol. 2003;24(5):258–70.
Romanowicz L, Galewska Z. Extracellular matrix remodeling of the umbilical cord in pre-eclampsia as a risk factor for fetal hypertension. J Pregnancy. 2011;54(2):69–75.
Kawazu T, Nishino T, Obata Y. Production and degradation of extracellular matrix in reversible glomerular lesions in rat model of habu snake venom-induced glomerulonephritis. Med Mol Morphol. 2012;45(4):190–8.
Mayer AM, Guzman M, Peksa R, et al. Differential effects of domoic acid and E. coli lipopolysaccharide on tumor necrosis factor-alpha, transforming growth factor-beta1 and matrix metalloproteinase-9 release by rat neonatal microglia: evaluation of the direct activation hypothesis. Mar Drugs. 2007;5(3):113–35.
Carvalho RR, Pellizzon CH, Justulin L Jr, et al. Effect of mangiferin on the development of periodontal disease: involvement of lipoxin A4, anti-chemotaxic action in leukocyte rolling. Chem Biol Interact. 2008;179(2):344–50.
Bellafiore M, Battaglia G, Bianco A, et al. The involvement of MMP-2 and MMP-9 in heart exercise-related angiogenesis. J Transl Med. 2013;11(1):283–9.
Cohen JN, Tewalt EF, Rouhani SJ, et al. Tolerogenic properties of lymphatic endothelial cells are controlled by the lymph node microenvironment. PLoS One. 2014;9(2):74–87.
Luo YM, Wan XH, Jiang DQ, et al. Effects of PPAR-gamma agonist rosiglitazone on MMP-9 and TIMP-1 expression of monocyte-derived macrophages isolated from patients with acute coronary syndrome. Zhonghua Xin Xue Guan Bing Za Zhi. 2009;37(8):739–45.
Kara S, Yildirim N, Ozer A, et al. Matrix metalloproteinase-2, tissue inhibitor of matrix metalloproteinase-2, and transforming growth factor beta 1 in the aqueous humor and serum of patients with pseudoexfoliation syndrome. Clin Ophthalmol. 2014;29(8):305–9.
Wang CL, Chen ZX, Li ZJ, et al. Effect of TIMP-2, MT1-MMP and MMP-2 expression on the in vitro invasive capacity of acute monocytic leukemia SHI-1 cells. Zhonghua Xue Ye Xue Za Zhi. 2010;31(12):798–803.
Groblewska M, Mroczko B, Kozlowski M, et al. Serum matrix metalloproteinase 2 and tissue inhibitor of matrix metalloproteinases 2 in esophageal cancer patients. Folia Histochem Cytobiol. 2012;50(4):590–8.
Wu TY, Zhang TH, Qu LM, et al. MiR-19a is correlated with prognosis and apoptosis of laryngeal squamous cell carcinoma by regulating TIMP-2 expression. Int J Clin Exp Pathol. 2013;7(1):56–63.
Groblewska M, Siewko M, Mroczko B, et al. The role of matrix metalloproteinases (MMPs) and their inhibitors (TIMPs) in the development of esophageal cancer. Folia Histochem Cytobiol. 2012;50(1):12–9.
Teijeira A, Rouzaut A, Melero I, et al. Initial afferent lymphatic vessels controlling outbound leukocyte traffic from skin to lymph nodes. Front Immunol. 2012;21(4):433–6.
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This work was supported by the National Natural Science Foundation of China (81272370).
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Yang, X., Zhai, N., Sun, M. et al. Influence of lymphatic endothelial cells on proliferation and invasiveness of esophageal carcinoma cells in vitro and lymphangiogenesis in vivo. Med Oncol 32, 222 (2015). https://doi.org/10.1007/s12032-015-0662-3
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DOI: https://doi.org/10.1007/s12032-015-0662-3