基于蛋白激酶NEK9/MTA2信号通路泛素化修饰探讨胃癌的转移机制  

Exploring the Metastatic Mechanism of Gastric Cancer Based on the Ubiquitination Modification of the Protein Kinase NEK9/MTA2 Signaling Pathway

作　　者：王志英 解祥军[2] 蔡珂[3] WANG Zhiying(Qingdao University,Shandong Qingdao 266000,China)

机构地区：[1]青岛大学,山东青岛266000 [2]山东省青岛市市立医院消化内科,山东青岛266000 [3]青岛大学医学院松山医院全科,山东青岛266000

出　　处：《河北医学》2024年第7期1087-1093,共7页Hebei Medicine

基　　金：山东省卫生健康科研基金项目,(编号:20220165)。

摘　　要：目的:基于永离有丝分裂基因A相关激酶9(NEK9)/转移相关肿瘤基因家族2(MTA2)信号通路泛素化修饰探讨胃癌(GC)的转移机制。方法:使用癌症基因组图谱(TCGA)和Kaplan-Meier Plotter数据库分析NEK9表达与GC分期、预后之间的关联。体外实验中,将GC细胞分为:对照组、shNC组、shNEK9组、shNC+NC-OE组、shNEK9+NC-OE组和shNEK9+MTA2-OE组。分别采用MTT和Transwell法测定细胞的增殖、迁移和侵袭,并通过Western blot检测NEK9、MTA2、上皮间充质转化(EMT)标记和PI3K/AKT信号通路蛋白表达。结果:TCGA数据库分析显示,NEK9 mRNA在肿瘤组织中的表达明显上调,并且与TNM分期较晚和NEK9高表达者的预后较差密切相关。此外,NEK9在7个GC细胞系中的表达明显高于正常胃上皮GES-1细胞(P<0.05)。与对照组相比,shNEK9组细胞活力、相对集落形成、EdU阳性细胞数、侵袭和迁移细胞数均显著降低(P<0.05)。此外,在shNEK9组细胞中,E-钙粘蛋白水平上调(P<0.05),波形蛋白水平下调(P<0.05)。通过免疫共沉淀试验证明NEK9与MTA2有相互作用。NEK9敲低加速了HGC-27细胞中MTA2的降解,并且MTA2泛素化在NEK9沉默的细胞中增加。与shNEK9+NC-OE组组相比,shNEK9+MTA2-OE组相对集落形成、EdU阳性细胞数和迁移和侵袭数均显著增加(P<0.05)。结论:NEK9在GC中明显上调,其敲低在体外抑制GC细胞的生长和转移。NEK9可能通过去泛素化途径稳定MTA2进而激活PI3K-AKT信号通路来对GC细胞产生促癌影响。Objective:To explore the metastasis mechanism of gastric cancer(GC)based on ubiquitination modification of the never-in-mitosis gene A-related kinase 9(NEK9)/metastasis-related tumor gene family 2(MTA2)signaling pathway.Methods:The Cancer Genome Atlas(TCGA)and Kaplan-Meier Plotter databases were used to analyze the association between NEK9 expression and GC staging and prognosis.In vitro experiments,GC cells were divided into the following groups:control group,shNC group,shNEK9 group,shNC+NC-OE group,shNEK9+NC-OE group,and shNEK9+MTA2-OE group.MTT and Transwell assays were used to determine cell proliferation,migration,and invasion,respectively.Western blot was used to detect the expression of NEK9,MTA2,epithelial-mesenchymal transition(EMT)markers,and PI3K/AKT signaling pathway proteins.Results:TCGA database analysis showed that NEK9 mRNA expression was significantly upregulated in tumor tissues and was closely related to poor prognosis in patients with advanced TNM stage and high NEK9 expression.In addition,NEK9 expression was significantly higher in 7 GC cell lines compared to normal gastric epithelial GES-1 cells(P<0.05).Compared to the control group,shNEK9 group cells showed significantly decreased cell viability,relative colony formation,EdU-positive cell number,invasion,and migration(P<0.05).Moreover,E-cadherin levels were upregulated(P<0.05)and vimentin levels were downregulated(P<0.05)in shNEK9 group cells.Immunoprecipitation assay demonstrated that NEK9 interacted with MTA2.NEK9 knockdown accelerated the degradation of MTA2 in HGC-27 cells,and MTA2 ubiquitination was increased in NEK9-silenced cells.Compared to the shNEK9+NC-OE group,shNEK9+MTA2-OE group cells showed significantly increased relative colony formation,EdU-positive cell number,and migration and invasion(P<0.05).Conclusion:NEK9 is significantly upregulated in GC and its knockdown suppresses GC cell growth and metastasis in vitro.NEK9 may promote tumorigenesis in GC cells by stabilizing MTA2 through deubiquitination and activating the PI3K-A

关 键 词：胃癌 永离有丝分裂基因A相关激酶9 转移相关肿瘤基因家族2 泛素化修饰 

分 类 号：R735.2[医药卫生—肿瘤]