ZNF436在胃癌中的表达及调控WNT/β-catenin对胃癌细胞迁移及侵袭的影响

Expression of ZNF436 in gastric cancer and its effect on the migration and invasion of gastric cancer cells by WNT/β-catenin

目的探讨锌指蛋白ZNF436在人胃癌细胞系和组织中的相对表达,及ZNF436对胃癌细胞MKN-28迁移和侵袭的影响和机制。方法以人胃癌细胞系,胃癌、配对癌旁组织和正常人胃组织为研究对象;实时定量PCR和免疫组织化学法分析ZNF436在胃癌细胞系,胃癌、配对癌旁组织和正常人胃组织中的相对表达;在线数据分析ZNF436的表达与预后关系;ZNF436-siRNA(实验组)、NC-siRNA(对照组)质粒转染人胃癌MKN028细胞,转染后48 h实时定量PCR验证ZNF436的敲降效率,Transwell实验分析敲降ZNF436后对实验组细胞迁移和侵袭影响;裸鼠转移瘤实验验证ZNF436体内对细胞转移的影响;免疫蛋白印迹实验验证ZNF436对WNT/β-catenin信号通路的影响。结果与正常胃上皮细胞系相比,ZNF436在人胃癌细胞系中高表达( P <0.001);同时与癌旁组织和正常人胃组织相比,ZNF436在胃癌组织中高表达( P <0.001);在线数据发现ZNF436的表达与胃癌的预后有关( P =0.0028);ZNF436基因表达越高,患者预后越差( P <0.001);与对照组相比,实验组细胞的迁移和侵袭能力明显受到抑制( P <0.001);实验组中裸鼠转移瘤的瘤大小和体积明显低于实验组( P < 0.001);低表达ZNF436后,WNT/β-catenin信号通路及其下游细胞因子明显受到抑制。结论ZNF436作为潜在癌基因在人胃癌组织中表达上调,通过促进WNT/β-catenin信号通路参与胃癌的进程。

ObjectiveThe aim of this study was to investigate the relative expression of zinc finger protein ZNF436 in human gastric cancer cells and tissues and its mechanism of ZNF436 on migration and invasion of gastric cancer MKN-28 cells. Methods RT-qPCR and immunohistochemistry were used to determine the expression of ZNF436 at the levels of mRNA and protein in human gastric cancer cells,gastric cancer and matched paracancerous tissues,and normal human gastric tissues.ZNF436 expression and prognostics were analyzed through online data.The ZNF436-siRNA(experimental group)and NC-siRNA(control group)plasmids were transfected into MKN028 cells.RT-qPCR was used to confirm the knockout efficiency of ZNF436 after transfection for 48 hours.The transwell assay was used to analyze the effect of ZNF436 on cell migration and invasion after knockout ZNF436.The effect of ZNF436 on cell metastasis was verified by nude mice metastasis experiments.The effect of ZNF436 on WNT/β-catenin signaling pathway was verified by immunoblotting.ResultsCompared with normal gastric epithelial cells,ZNF436 was highly expressed in human gastric cancer cells( P <0.001).ZNF436 was highly expressed in gastric cancer tissues compared with adjacent normal tissues and normal human gastric tissues( P <0.001).The expression of ZNF436 was associated with the prognosis of gastric cancer from online data( P =0.0028);the high expression of ZNF436 gene,the worse prognosis of patients( P < 0.001);When compared with the control group,the migration and invasion ability in the experimental group was significantly inhibited( P <0.001);the size and volume of metastasis in nude mice in the experimental group were significantly lower than those in the experimental group( P <0.001).After the low expression of ZNF436,the WNT/β-catenin signaling pathway and its downstream cytokines were significantly inhibited.ConclusionZNF436 is up-regulated as a potential oncogene in human gastric cancer and participates in the progression of gastric cancer by promoting WNT/β-catenin signaling pathway.