去泛素化酶USP14通过稳定β-catenin蛋白促进子宫内膜癌细胞增殖和上皮间充质转化  

Deubiquitinating enzyme USP14 promotes endometrial cancer cell proliferation and epithelial mesenchymal transformation via stabilizingβ-catenin protein

作　　者：张易欣 刘平[1] 王菊荣[1] 李美艳[1] 杨晓明[1] 姜黎黎 张强[2] ZHANG Yixin;LIU Ping;WANG Jurong;LI Meiyan;YANG Xiaoming;JIANG Lili;ZHANG Qiang(Department of Obstetrics and Gynecology,Handan Central Hospital,Handan 056000,China;Department of Ultrasound Medicine,Handan Central Hospital,Handan 056000,China)

机构地区：[1]邯郸市中心医院妇一科,056000 [2]邯郸市中心医院超声医学科,056000

出　　处：《免疫学杂志》2024年第6期533-538,共6页Immunological Journal

基　　金：邯郸市科学技术研究与发展计划(1823208055ZC)。

摘　　要：目的探究去泛素化酶USP14和β-连环蛋白(β-catenin)在子宫内膜癌(EC)中的生物学功能和相互作用机制。方法通过实时荧光定量PCR和免疫组织化学检测EC组织和癌旁健康组织中USP14和β-catenin的表达;通过蛋白质免疫印迹和蛋白质免疫沉淀的方法,在EC来源的HEA-1C和Ishikawa细胞系中探讨USP14对β-catenin蛋白质稳定性和泛素化调节的作用;通过细胞增殖试验(CCK-8)验证USP14和β-catenin对EC细胞增殖的作用;通过质粒转染验证USP14对β-catenin的调节作用;通过蛋白质免疫印迹验证USP14和β-catenin对EC细胞上皮-间充质转化(EMT)相关蛋白表达的调控。结果USP14和β-catenin在EC组织中的表达高于癌旁健康组织,USP14与β-catenin在EC细胞系中存在相互作用,USP14通过其去泛素化活性维持β-catenin的蛋白稳定性,用USP14特异性抑制剂处理后能够增强β-catenin的泛素化水平和下调β-catenin表达。用USP14特异性抑制剂能抑制EC细胞增殖和EMT相关蛋白的表达,并且过表达β-catenin能逆转USP14抑制剂导致的EC细胞生长抑制和EMT抑制。结论USP14在EC细胞中通过调节β-catenin的泛素化水平维持其蛋白稳定性,其对细胞增殖和EMT的调控需要β-catenin参与。This study was designed to investigate the biological function and interaction mechanism of deubiquitinating enzyme USP14 andβ-catenin in endometrial carcinoma(EC).The expression of USP14 andβ-catenin in EC tissues and adjacent healthy tissues were detected by real-time fluorescence quantitative PCR and immunohistochemistry,while the effects of USP14 onβ-catenin protein stability and ubiquitination were investigated by Western blotting and protein immunoprecipitation using HEA-1C and Ishikawa cell lines derived from EC.The effect of USP14 andβ-catenin on EC cell proliferation was verified by cell proliferation assay(CCK-8);the regulatory effect of USP14 onβ-catenin was verified by plasmid transfection.The regulation of USP14 andβ-catenin on the expression of proteins related to epithelial-mesenchymal transition(EMT)in EC cells were verified by Western blotting.Data showed that the expression of USP14 andβ-catenin in EC tissues were higher than those in adjacent healthy tissues,and USP14 andβ-catenin interacted in EC cell line.USP14 maintained the protein stability ofβ-catenin through its deubiquitination activity.USP14-specific inhibitors could enhance the ubiquitination level ofβ-catenin and down-regulate the expression ofβ-catenin.USP14-specific inhibitors also inhibited EC cell proliferation and inhibited the expression of EMT-related proteins,while overexpression ofβ-catenin reversed the inhibition of EC cell growth and EMT induced by USP14 inhibitors.In conclusion,USP14 maintainsβ-catenin protein stability in EC cells by regulating the ubiquitination level,and the regulation of cell proliferation and EMT requiresβ-catenin.

关 键 词：子宫内膜癌 上皮间充质转化 去泛素化活性 蛋白稳定性 

分 类 号：R737.33[医药卫生—肿瘤]