摘要
目的:探讨表没食子儿茶素没食子酸酯(Epigallocatechin-3-gallate,EGCG)体外抑制潜伏性膜蛋白1转化鼻咽上皮细胞NP69(NP69-LMP1)增殖的机制。方法:采用MTT法检测EGCG抑制NP69-LMP1细胞增殖的IC50值;选用细胞生长曲线、软琼脂集落形成实验和平皿克隆形成实验观察EGCG抑制NP69-LMP1细胞增殖的作用;采用Western-blot检测Wnt1、β-catenin的表达及磷酸化。结果:EGCG抑制NP69-LMP1细胞增殖的IC50值为47.7mg.L-1;EGCG对NP69-LMP1细胞增殖的抑制作用呈浓度和时间依赖性(n=3,P<0.05)。EGCG呈浓度依赖性抑制Wnt1和β-catenin蛋白的表达,促进β-catenin蛋白的磷酸化。结论:EGCG通过降低Wnt1和β-catenin蛋白表达,增加β-catenin蛋白磷酸化水平,发挥对NP69-LMP1细胞增殖的抑制作用。
Objective：This study aimed to the investigate mechanism of Epigallocatechin-3-gallate（EGCG） inhibited the Proliferation of NP69-LMP1 cells in vitro.Methods：The 50 % inhibiting concentration（IC50） of EGCG-inhibited proliferation of NP69-LMP1 cells was detected by MTT method.EGCG-inhibited proliferation of NP69-LMP1 cells was observed through the cellular growth curve,soft agar formation and colony formation.The expression of Wnt1,β-catenin proteins and its phosphorylation level in NP69-LMP1 cells treated by EGCG were analyzed by Western-blot.Results：IC50 of EGCG-inhibited proliferation of NP69-LMP1 cells was 47.7 mg.L-1.The proliferation of NP69-LMP1 cell was inhibited along with EGCG concentration increasing and treatment time extending（n=3,P0.05）.The expression of Wnt1 and β-catenin proteins in NP69-LMP1 cells was obviously degraded,but the phosphorylation level of β-catenin successively was raised with EGCG concentration increasing.Conclusion：EGCG decreased the expression of Wnt1 and β-catenin protein and increased the phosphorylation level of β-catenin protein to inhibit the proliferation of NP69-LMP1.
出处
《现代生物医学进展》
CAS
2012年第36期7034-7039,共6页
Progress in Modern Biomedicine
基金
湖南省高校创新平台开放基金(10K052)
湖南省研究生科研创新项目(CX2010B380)
湖南省大学生研究性学习和创新性实验计划项目(CXSY-SJ-09007
CXSY-SJ-10192)
湖南省教育厅科研项目(11C1112)
衡阳市科技局科研项目(2011kj52
2011kj53)
