过氧蛋白同源物通过调控Wnt/β-连环蛋白信号通路影响胃癌MGC803细胞生长和侵袭迁移能力  

Effects of peroxidasin homolog on growth,invasion and migration of gastric cancer MGC803 cells by regulating Wnt/beta-catenin signaling pathway

作　　者：钟文进 林建安[1] 孙亚锋[1] 许建华[1] 叶凯[1] Zhong Wenjing;Lin Jianan;Sun Yafeng;Xu Jianhua;Ye Kai(Department of Gastrointestinal Surgery,the Second Affiliated Hospital of Fujian Medical University,Quanzhou 362002,China)

机构地区：[1]福建医科大学附属第二医院胃肠外科,泉州362002

出　　处：《中华实验外科杂志》2020年第3期412-415,共4页Chinese Journal of Experimental Surgery

摘　　要：目的探讨过氧蛋白同源物(PXDN)基因对人胃癌MGC803细胞生长和侵袭迁移能力的影响及其机制。方法收集从2019年4月到2019年6月期间福建医科大学附属第二医院胃肠外科确诊为胃癌的未经过放疗和化疗的手术标本及配对的癌旁组织,各10例。通过癌症肿瘤基因组图谱(TCGA)数据库筛选与胃癌相关的突变基因,并通过筛选GEO数据库中与胃癌相关的RNA测序数据集(GSE122796)分析突变基因的差异表达状况。收集临床上未经放、化疗的胃癌组织及配对的癌旁组织各10例,采用实时定量反转录聚合酶链反应(RT-qPCR)法和蛋白质印迹法(Western blot)检测PXDN在癌旁组织和胃癌组织中的表达。通过Western blot检测PXDN在4种人胃癌细胞株的表达。构建小干扰RNA沉默PXDN的表达,将培养的细胞随机随机分为NC-SiRNA组(转染阴性对照组)和PXDN-SiRNA组,并采用细胞计数试剂盒(CCK-8)、划痕实验、Transwell法,检测MGC803细胞的增殖、侵袭和迁移的变化。此外,Western blot检测Wnt/β-连环蛋白(β-catenin)信号通路的改变。应用SPSS 22.0统计软件分析,采用t检验或单因素方差进行分析。结果TCGA与GEO数据库显示PXDN是胃癌发病相关的高突变率(15.4%)及显著差异基因[Log2 FC=2.83±0.72,padj=0.031,P<0.05]。RT-qPCR和Western blot结果显示PXDN在胃癌组织表达量明显上调[(0.92±0.12)比(0.61±0.09)、(1.65±0.23)比(1.00±0.25),t=12.680、17.520,P<0.05],差异有统计学意义。Western blot检测PXDN在MGC803细胞株表达量较其他细胞株高(F=23.810,P<0.05),差异有统计学意义。沉默PXDN基因可显著抑制MGC803细胞的增殖[吸光度(A)值:(0.62±0.05)比(1.05±0.08),t=7.740,P<0.05]、迁移[(41.25±5.67)个比(85.23±4.28)个,t=21.120,P<0.05]、侵袭[(42.47±3.98)个比(86.16±4.19)个,t=20.090,P<0.05],差异有统计学意义。此外,沉默PXDN后,糖原合成酶激酶-3β(Gsk-3β)蛋白表达水平显著上调,β-连环蛋白(β-catenin)、细胞周期�Objective To investigate the effect of Peroxidasin Homolog(PXDN)on the growth,invasion and migration of human gastric cancer MGC803 cells and the underlying mechanism.Methods The mutant genes highly related to gastric cancer were screened through the cancer genome atlas(TCGA)database,and the differential expression levels of these genes were analyzed by high throughput gastric cancer RNA sequencing dataset screened in GEO database(GSE122796).Quantitative reverse transcription polymerase chain reaction(RT-qPCR)and Western blotting were used to detect the expression of PXDN in paracancer and gastric cancer tissues respectively.The expression levels of PXDN in four human gastric cancer cell lines were detected by Western blotting.PXDN was silenced by small interference RNA,Cell line MGC803 was randomly divided into two groups-NC-siRNA and PXDN-siRNA,and cell counting kit-8(CCK-8)assay,scratch test and Transwell method were used to detect the proliferation,invasion and migration of MGC803 cells.In addition,Western immunoblotting was used to detect the variations in the Wnt/β-catenin signaling pathway.Results TCGA and GEO database showed that PXDN was a highly mutated(15.4%)and expressed gene associated with gastric cancer(Log2 FC=2.83±0.72,padj=0.031,P<0.05).The results of RT-qPCR and Western immunoblotting showed that the expression of PXDN in gastric cancer tissues was significantly up-regulated(0.92±0.12 vs.0.61±0.09;1.65±0.23 vs.1.0±0.25,t=12.680,17.520,P<0.05).The expression of PXDN in MGC803 cells was significantly higher than that in other cell lines by Western blotting(F=23.810,P<0.05).PXDN silencing significantly inhibited the proliferation[optical density(A)value:0.62±0.05 vs.1.05±0.08,t=7.740,P<0.05],migration(41.25±5.67 vs.85.23±4.28,t=21.120,P<0.05)and invasion(42.47±3.98 vs.86.16±4.19,t=20.090,P<0.05)of MGC803 cells.In addition,the silencing of PXDN significantly up-regulated the expression level of GSK-3βand significantly down-regulated the protein levels ofβ-catenin and Cyclin D1(t=12.59

关 键 词：胃癌 过氧蛋白同源物 增殖 迁移和侵袭 Wnt3a/β-连环蛋白 

分 类 号：R735.2[医药卫生—肿瘤]