脯氨酰羟化酶3通过β-catenin/TCF通路抑制胃癌细胞的克隆形成和迁移  

PHD3 inhibits the clone formation and migration of gastric cancer cells by β-catenin/TCF signaling

作　　者：陈吉祥[1] 蒋国雄[1] 党胜春[1] 瞿建国[1] 谢嵘[1] 崔磊[1] 

机构地区：[1]江苏大学附属医院普外科,江苏镇江212001

出　　处：《江苏大学学报（医学版）》2017年第3期233-238,共6页Journal of Jiangsu University：Medicine Edition

基　　金：镇江市社会发展项目(SH2015066);江苏省"六大人才高峰"资助项目(2016-WSN-007)

摘　　要：目的:探讨脯氨酰羟化酶(prolyl hydroxylase,PHD)3对胃癌细胞克隆形成和迁移能力的影响及可能的分子机制。方法:运用细胞转染上调和沉默AGS和BGC823胃癌细胞系中PHD3的表达;在转染后的细胞系中,通过软琼脂实验和迁移实验检测克隆形成和迁移能力的变化,采用蛋白质印迹法检测细胞周期蛋白D1(Cyclin D1)、cMyc及Twist的表达;在AGS和BGC823细胞系中,用免疫荧光及免疫共沉淀检测PHD3和Dishevelled-2(DVL2)的相互作用。结果:上调PHD3后,AGS和BGC823克隆形成及迁移能力明显降低(均P<0.01);沉默PHD3后,AGS和BGC823克隆形成及迁移能力明显增强(均P<0.01);上调PHD3可抑制转染细胞系中Cyclin D1、c-Myc及Twist的表达,沉默PHD3可促进Cyclin D1、c-Myc及Twist的表达;PHD3与DVL2在胃癌细胞中存在相互作用。结论:PHD3可能通过β-catenin/TCF信号通路抑制胃癌细胞的克隆形成和迁移能力。Objective：To explore the influence and the possible molecular mechanism of the prolyl hydroxylase（PHD）3 on clone formation and migration of gastric cancer cells.Methods：The level of PHD3 in AGS and BGC823 cell lines was up-regulated and silenced by cell transfection with overexpression plasmid or siRNA.The soft agar assay and chamber assay were used to detect the change of clone formation and migration in the gastric cancer cells.The level of Cyclin D1,c-Myc and Twist in the gastric cancer cells were measured with Western blotting.Immunofluorescence and Co-immunoprecipitation assays were used to determine the interaction between PHD3 and Dishevelled-2（DVL2）.Results：After over-expressing of PHD3,the ability of clone formation and migration of AGS and BGC823 were significantly attenuated（P〈0.01）.Conversely, they were significantly enhanced after silencing PHD3 （P〈0.01）.Over-expression of PHD3 inhibited the level of Cyclin D1,c-Myc and Twist, down-regulated of PHD3 elevated the level of Cyclin D1,c-Myc and Twist.PHD3 and DVL2 have interaction in gastric cancer cells.Conclusion：PHD3 can restrain the clone formation and migration of gastric cancer cells by modification of β-catenin/TCF signaling.

关 键 词：胃癌 脯氨酰羟化酶3 β-catenin/TCF通路 克隆形成和迁移 

分 类 号：R73-362[医药卫生—肿瘤]