表没食子儿茶素没食子酸酯对牙源性角化囊肿上皮细胞增殖和凋亡的影响  

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

作　　者：杨绍滨 薛娇 庞宝兴[1,3] 袁荣涛[1,2] YANG Shao-bin;XUE Jiao;PANG Bao-xing;YUAN Rong-tao(College of Stomatology,Qingdao University.Qingdao 266000;Center of Oral Medicine,Qingdao Municipal Hospital.Qingdao 266011;Department of Oral and MaxiUofacial Surgery,The Affiliated Hospital of Qingdao University.Qingdao 266001,Shandong Province,China)

机构地区：[1]青岛大学口腔医学院,山东青岛266000 [2]青岛大学附属青岛市立医院口腔医学中心,山东青岛266071 [3]青岛大学附属医院口腔颌面外科,山东青岛266001

出　　处：《中国口腔颌面外科杂志》2018年第6期481-487,共7页China Journal of Oral and Maxillofacial Surgery

基　　金：山东省自然科学基金(ZR2016HM34)

摘　　要：目的:探讨表没食子儿茶素没食子酸酯(EGCG)对牙源性角化囊肿(OKC)上皮细胞增殖、凋亡的影响,以及与Wnt信号通路相关基因FZD3和JNK3表达的关系。方法:采用原代培养的人牙源性角化囊肿上皮细胞,分别用不同浓度的EGCG处理24、48、72 h,CCK-8法检测细胞增殖,FITC-Annexin V/PI染色法检测细胞凋亡,流式细胞术检测细胞周期分布,RT-PCR、Western免疫印迹检测Wnt信号通路相关基因FZD3和JNK3的表达。采用SPSS 22.0软件包对数据进行统计学分析。结果:EGCG以剂量-时间依赖方式抑制OKC上皮细胞增殖。低浓度EGCG(0～20μmol/L)对细胞增殖无显著影响(P>0.05),高浓度EGCG(40～320μmol/L)对细胞增殖有显著抑制作用(P<0.05),且随着药物浓度增加及作用时间延长,抑制作用逐渐增强(P<0.05)。EGCG (20、160、320μmol/L)诱导OKC上皮细胞凋亡,呈剂量依赖性(P<0.05),并将细胞周期阻滞于G1期。EGCG可下调Wnt信号通路相关基因FZD3和JNK3的表达(P<0.05)。结论:EGCG抑制OKC上皮细胞增殖,诱导细胞凋亡,其机制可能与阻断Wnt信号通路相关基因FZD3和JNK3的表达有关。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.

关 键 词：表没食子儿茶素没食子酸酯 牙源性角化囊肿 增殖 凋亡 卷曲蛋白受体3 c—Jun氨基末端激酶3 

分 类 号：R739.8[医药卫生—肿瘤]