摘要
目的:探究肿瘤坏死因子-α诱导蛋白8样因子2(TIPE2)是否通过抑制Wnt/β-catenin信号通路影响胃癌(GC)细胞增殖、迁移和上皮-间质转化(EMT)。方法:qRT-PCR和Western blot检测不同细胞中TIPE2 mRNA和蛋白表达。体外培养MKN45细胞,依次分为:Control组(不做任何处理)、AD-EV组(转染AD-EV)、AD-TIPE2组(转染AD-TIPE2)、LiCl组(Wnt/β-catenin信号通路激活剂LiCl处理)、AD-TIPE2+LiCl组(转染AD-TIPE2后LiCl处理)。MTT和集落形成实验检测细胞增殖;Transwell检测细胞迁移;Western blot检测细胞TIPE2、E-cadherin、N-cadherin、Wnt2、β-catenin和Twist蛋白表达;免疫荧光染色检测细胞TIPE2、E-cadherin及N-cadherin表达。结果:GC细胞株中TIPE2表达显著下调,且在MKN45细胞中表达最低。与AD-EV组相比,AD-TIPE2组MKN45细胞增殖活力、集落形成数、迁移细胞数及N-cadherin、TIPE2、Wnt2、β-catenin、Twist、c-Myc和CyclinD1蛋白表达均显著降低,TIPE2和E-cadherin表达显著增高(P<0.05);与AD-TIPE2组相比,AD-TIPE2+LiCl组MKN45细胞增殖活力、集落形成数、迁移细胞数及N-cadherin、TIPE2、Wnt2、β-catenin、Twist、c-Myc和CyclinD1蛋白表达均显著升高,TIPE2和E-cadherin表达显著降低(P<0.05);与LiCl组相比,AD-TIPE2+LiCl组MKN45细胞增殖活力、集落形成数、迁移细胞数及N-cadherin、TIPE2、Wnt2、β-catenin、Twist、c-Myc和CyclinD1蛋白表达均显著降低,TIPE2和E-cadherin表达显著升高(P<0.05)。结论:上调TIPE2可通过抑制Wnt/β-catenin信号通路抑制GC细胞增殖、迁移和EMT。
Objective:To explore effects of tumor necrosis factor-α-induced protein-8 like-2(TIPE2)on proliferation,migration and epithelial-mesenchymal transition(EMT)of gastric cancer(GC)cells through Wnt/β-catenin signaling pathway.Methods:qRT-PCR and Western blot were used to detect TIPE2 mRNA and protein expressions in different cell types.MKN45 cells were cultured in vitro and divided into:Control group(without any treatment),AD-EV group(transfected with AD-EV),AD-TIPE2 group(transfected with AD-TIPE2),LiCl group(treated with Wnt/β-catenin signaling pathway activator,LiCl)and AD-TIPE2+LiCl group(treated with LiCl after transfection of AD-TIPE2).MTT and colony formation test were used to detect cells proliferation,Transwell was used to detect cells migration;Western blot was used to detect expressions of TIPE2,E-cadherin,N-cadherin,Wnt2,β-catenin and Twist proteins of cells;immunofluorescence staining was used to detect expressions of TIPE2,E-cadherin and N-cadherin of cells.Results:TIPE2 expression was significantly down-regulated in GC cell lines,and was the lowest in MKN45 cells.Compared with AD-EV group,MKN45 cell proliferation activity,number of colonies formed,number of migrating cells and expressions of N-cadherin,TIPE2,Wnt2,β-catenin,Twist,c-Myc and CyclinD1 proteins in AD-TIPE2 group were significantly decreased,and expressions of TIPE2 and E-cadherin were significantly increased(P<0.05);compared with AD-TIPE2 group,MKN45 cell proliferation activity,number of colonies formed,number of migrating cells and expressions of N-cadherin,TIPE2,Wnt2,β-catenin,Twist,c-Myc and CyclinD1 proteins in AD-TIPE2+LiCl group were significantly increased,and expressions of TIPE2 and E-cadherin were significantly decreased(P<0.05);compared with LiCl group,MKN45 cell proliferation activity,number of colonies formed,number of migrating cells and expressions of N-cadherin,TIPE2,Wnt2,β-catenin,Twist,c-Myc and CyclinD1 proteins in AD-TIPE2+LiCl group were significantly decreased,and expressions of TIPE2 and E-cadherin were significantly increased(P<0.05).Conclusion:Up-regulation of TIPE2 can inhibit proliferation,migration and EMT of GC cells by inhibiting Wnt/β-catenin signaling pathway.
作者
王燕
张宝
王交莉
WANG Yan;ZHANG Bao;WANG Jiaoli(Department of Oncology,Jinan Hospital of Integrated Traditional Chinese and Western Medicine,Jinan 271100,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2023年第8期1700-1705,共6页
Chinese Journal of Immunology
