Effect of epigallocatechin-3-gallate on proliferation and apoptosis of human odontogenic keratocyst epithelial cells

doi:10.19438/j.cjoms.2018.06.001

Abstract

Abstract: PURPOSE: To investigate the effects of EGCG on proliferation, apoptosis and Wnt pathway-related genes, FZD3 and JNK3, of human odontogenic keratocyst (OKC) epithelial cells in vitro. METHODS: OKC epithelial cells isolated from human odontogenic keratocyst tissue were primarily cultured. Cells were treated with different concentrations of EGCG (0, 20, 40, 80, 160, 320 μmol/L) for 24, 48 and 72 h. The proliferation of OKC epithelial cells was evaluated by CCK-8 assay. Cell apoptosis and cell cycle distribution were detected by flow cytometry. mRNA expression of Wnt signaling pathway correlated genes, FZD3 and JNK3, was determined by RT-PCR. The proteins expression was determined by Western blotting. Statistical analysis was performed using SPSS 22.0 software package. RESULTS: EGCG inhibited cell proliferation in a dose and time-dependent pattern. Low-concentration EGCG (0-20 μmol/L) had no obvious effect on OKC epithelial cells (P>0.05), while high concentrations of EGCG (40-320 μmol/L) had suppressive effect on OKC epithelial cells (P<0.05). With increasing concentration and prolonged action time, the inhibition rate increased (P<0.05). EGCG (20, 160, 320μmol/L) induced apoptosis of cells (P<0.05), and cell cycle was arrested in G1 phase. In addition, both mRNA and protein expression of Wnt pathway related genes, FZD3 and JNK3, was down-regulated significantly (P<0.05). CONCLUSIONS: EGCG inhibits proliferation, induces apoptosis and arrests cell cycle in G1 phase of OKC cells by a possible mechanism of blocking Wnt signaling pathway.

Key words: Epigallocatechin-3-gallate, Odontogenic keratocyst, Proliferation, Apoptosis, Frizzled receptor 3, c-Jun N-terminal kinase 3

CLC Number:

R739.8

YANG Shao-bin, XUE Jiao, PANG Bao-xing, YUAN Rong-tao. Effect of epigallocatechin-3-gallate on proliferation and apoptosis of human odontogenic keratocyst epithelial cells[J]. China Journal of Oral and Maxillofacial Surgery, 2018, 16(6): 481-487.

References

[1] Speight PM, Takata T.New tumour entities in the 4th edition of the World Health Organization Classification of Head and Neck tumours: odontogenic and maxillofacial bone tumours[J]. Virchows Arch, 2018, 472(3): 331-339.
[2] Mendes RA, Carvalho JF, van der Waal I. Characterization and management of the keratocystic odontogenic tumor in relation to its histopathological and biological features[J]. Oral Oncol, 2010, 46(4): 219-225.
[3] Singh BN, Shankar S, Srivastava RK.Green tea catechin, epigallocatechin-3-gallate (EGCG): mechanisms, perspectives and clinical applications[J]. Biochem Pharmacol, 2011, 82(12): 1807-1821.
[4] Khan N, Afaq F, Saleem M, et al.Targeting multiple signaling pathways by green tea polyphenol (-)-epigallocatechin-3-gallate[J]. Cancer Res, 2006, 66(5): 2500-2505.
[5] Johnson NR, Batstone MD, Savage NW.Management and recurrence of keratocystic odontogenic tumor: a systematic review[J]. Oral Surg Oral Med Oral Pathol Oral Radiol, 2013, 116(4): e271-e276.
[6] 胡永杰, 李思毅, 徐立群, 等. 减压术治疗下颌骨大型牙源性角化囊肿的临床研究[J]. 中国口腔颌面外科杂志, 2005, 3(4): 299-302.
[7] Awni S, Conn B.Decompression of keratocystic odontogenic tumors leading to increased fibrosis, but without any change in epithelial proliferation[J]. Oral Surg Oral Med Oral Pathol Oral Radiol, 2017, 123(6): 634-644.
[8] 胡秀莲, 王兴, 林野, 等. 人口腔黏膜角化上皮细胞的体外培养及生物学特性研究[J]. 中国口腔颌面外科杂志, 2004, 2(3): 173-176.
[9] Guimaraes DM, Antunes DM, Saturno JL, et al.Immunohistochemical expression of WNT5A and MMPs in odontogenic epithelial tumors and cysts[J]. Acta Histochem, 2015, 117(8): 667-674.
[10] Hakim SG, Kosmehl H, Sieg P, et al.Altered expression of cell-cell adhesion molecules beta-catenin/E-cadherin and related Wnt-signaling pathway in sporadic and syndromal keratocystic odontogenic tumors[J]. Clin Oral Investig, 2011, 15(3): 321-328.
[11] 孔丽, 王炳超, 袁荣涛, 等. Wnt信号通路相关基因在颌骨牙源性角化囊性瘤中的表达及意义[J]. 上海口腔医学, 2015, 24(1): 89-93.
[12] Chen PN, Chu SC, Kuo WH, et al.Epigallocatechin-3 gallate inhibits invasion, epithelial-mesenchymal transition, and tumor growth in oral cancer cells[J]. J Agric Food Chem, 2011, 59(8): 3836-3844.
[13] Xiang LP, Wang A, Ye JH, et al.Suppressive effects of tea catechins on breast cancer[J]. Nutrients, 2016, 8(8): E458.
[14] Sakamoto Y, Terashita N, Muraguchi T, et al.Effects of epigallocatechin-3-gallate (EGCG) on A549 lung cancer tumor growth and angiogenesis[J]. Biosci Biotechnol Biochem, 2013, 77(9): 1799-1803.
[15] Yang C, Du W, Yang D.Inhibition of green tea polyphenol EGCG[(-)-epigallocatechin-3-gallate] on the proliferation of gastric cancer cells by suppressing canonical wnt/beta-catenin signalling pathway[J]. Int J Food Sci Nutr, 2016, 67(7): 818-827.
[16] Min KJ, Kwon TK.Anticancer effects and molecular mechanisms of epigallocatechin-3-gallate[J]. Integr Med Res, 2014, 3(1): 16-24.
[17] Kikuchi A, Yamamoto H, Sato A.Selective activation mechanisms of Wnt signaling pathways[J]. Trends Cell Biol, 2009, 19(3): 119-129.
[18] Katoh M.WNT/PCP signaling pathway and human cancer (review)[J]. Oncol Rep, 2005, 14(6): 1583-1588.
[19] Görögh T, Bèress L, Quabius ES, et al.Head and neck cancer cells and xenografts are very sensitive to palytoxin: decrease of c-jun n-terminale kinase-3 expression enhances palytoxin toxicity[J]. Mol Cancer, 2013, 12: 12.
[20] Long J, Cai L, Li J, et al.JNK3 involvement in nerve cell apoptosis and neurofunctional recovery after traumatic brain injury[J]. Neural Regen Res, 2013, 8(16):1491-1499.
[21] Pirianov G, Brywe KG, Mallard C, et al.Deletion of the c-Jun N-terminal kinase 3 gene protects neonatal mice against cerebral hypoxic-ischaemic injury[J]. J Cereb Blood Flow Metab, 2007, 27(5): 1022-1032.