三叶因子3在甲状腺乳头状癌TPC-1细胞株黏附中的作用机制  

Mechanism of trefoil factor 3 in adhesion of papillary thyroid carcinoma TPC-1 cell

作　　者：林旭[1] 代金 吴靖芳[1] 张静[1] 张文静[1] 薛刚[2] 高福禄[3] Lin Xu;Dai Jin;Wu Jingfang;Zhang Jing;Zhang Wenjing;Xue Gang;Gao Fulu(Basic Medical College,Head and Neck Surgery,First Affiliated Hospital, Hebei North University,Zhangjiakou 075000;Department of Otolaryngology,Head and Neck Surgery,First Affiliated Hospital, Hebei North University,Zhangjiakou 075000;Hebei Normal University,Shijiazhuang 050024,China)

机构地区：[1]河北北方学院基础医学院,张家口075000 [2]河北北方学院附属第一医院耳鼻咽喉头颈外科,张家口075000 [3]河北师范大学,石家庄050024

出　　处：《解剖学杂志》2018年第5期505-509,共5页Chinese Journal of Anatomy

基　　金：张家口市科技攻关计划(No:1411043H;1611055H)

摘　　要：目的:探讨沉默三叶因子3(TFF3)表达对甲状腺乳头状癌(PTC)TPC-1细胞黏附作用的影响。方法:稳定敲低TFF3表达的TPC-1细胞株(shRNA-TFF3组)、转染空载体的TPC-1细胞株(shRNAC组)、TPC-1细胞(TPC-1组)分别进行同质黏附实验、异质黏附实验;qPCR检测TFF3mRNA及WNT/β-catenin信号通路关键分子β-catenin mRNA水平,免疫印迹和免疫细胞化学检测β-catenin、E-cadherin、ALCAM、MMP-9、MMP-2蛋白表达水平。结果:细胞同质黏附实验显示shRNA-TFF3组黏附细胞数显著高于shRNAC组和TPC-1组。细胞异质实验显示shRNA-TFF3组黏附细胞数显著低于shRNAC组和TPC-1组,shRNAC组和TPC-1组同质和异质黏附数差异无统计学意义。qPCR结果显示shRNA-TFF3组TFF3mRNA、β-catenin mRNA水平显著低于shRNAC组和TPC-1组,shRNAC组和TPC-1组TFF3 mRNA、β-catenin mRNA水平差异无统计学意义。免疫细胞化学与免疫印迹结果显示沉默TFF3后ALCAM、MMP-9、MMP-2、β-catenin蛋白表达水平显著降低;E-cadherin蛋白表达水平显著升高,shRNAC组和TPC-1组差异无统计学意义。结论:敲低TFF3可能通过WNT/β-catenin信号通路影响PTC细胞TPC-1的黏附能力。Objective:To explore the effect of silencing trefoil factor 3(TFF3)expression on adhesion of papillary thyroid carcinoma TPC-1 cells.Methods:Homotyptic adhesion and heterotyptic adhesion experiment were used to assess the adhesive ability among stable knockdown TFF3(shRNA-TFF3)-TPC-1 cells,scrambled shRNA control(shRNAC)-TPC-1 cells and blank TPC-1 cells.WNT/β-catenin and TFF3 mRNA level was detected by quantitative RT-PCR(qRT-PCR).The protein expression levels of MMP-9,MMP-2,ALCAM,β-catenin and E-cadherin were detected by Western blotting and immunocytochemistry(ICC)technique.Results:Knockdown of TFF3 significantly increased the number of homotyptic adhesion compared with shRNAC group and TPC-1 group.The result of cell-matrix adhesive texts suggested that the number of heterotyptic adhesion cells was smaller than that of shRNAC group and TPC-1 group after inhibiting the expression of TFF3.There was no significant difference between shRNAC group and TPC-1 group in both homotyptic adhesion and heterotyptic adhesion test.Knockdown of TFF3 significantly decreased the mRNA level ofβ-catenin and TFF3,and inhibited the protein expression level of ALCAM,MMP-9,MMP-2,β-catenin,but it increased the protein expression level of E-cadherin.Conclusion:Siliencing TFF3 expression could affect the ability of adhesion in TPC-1 cells via the WNT/β-catenin signalling pathway.

关 键 词：甲状腺乳头状癌 黏附 三叶因子3 WNT/Β-CATENIN信号通路 

分 类 号：R736.1[医药卫生—肿瘤]