USP7-MDM2-p53信号轴对子宫内膜癌细胞增殖、凋亡和细胞周期的影响  

Impacts of USP7-MDM2-p53 signal axis on proliferation,apoptosis and cell cycle of endometrial carcinoma cells

作　　者：魏伟[1] 赵慧娟 刘湘翠 WEI Wei;ZHAO Huijuan;LIU Xiangcui(Department of Obstetrics and Gynecology,Heze Medical College,Shandong Heze 274000,China;Department of Gynecology,Affiliated Hospital of Heze Medical College,Shandong Heze 274000,China;Department of Gynecology,Peony People's Hospital of Heze,Shandong Heze 274000,China)

机构地区：[1]菏泽医学专科学校妇产科教研室,山东菏泽274000 [2]菏泽医专附属医院妇科,山东菏泽274000 [3]菏泽市牡丹人民医院妇科,山东菏泽274000

出　　处：《现代肿瘤医学》2024年第2期214-220,共7页Journal of Modern Oncology

基　　金：山东省卫生健康委员会资助项目(编号:WS2019339)。

摘　　要：目的:探讨泛素特异性蛋白酶7(USP7)调节Mdm2 p53结合蛋白同源物(MDM2)-p53轴对子宫内膜癌细胞增殖、凋亡和细胞周期的影响。方法:Western blot检测人子宫内膜癌组织、癌旁组织、人子宫内膜上皮细胞hEEC及人子宫内膜癌细胞系Ishikawa、HEC-1-A、KLE中USP7蛋白表达。将Ishikawa细胞分为NC组、P22077(USP7抑制剂)组、pcDNA组、pcDNA-MDM2组、P22077+pcDNA组、P22077+pcDNA-MDM2组,CCK-8法和克隆形成实验检测Ishikawa细胞增殖;流式细胞术检测Ishikawa细胞凋亡与细胞周期变化;Western blot检测Ishikawa细胞中USP7、细胞周期蛋白D1(CyclinD1)、周期素依赖性激酶2(CDK2)、Bcl-2相关X蛋白(Bax)、MDM2、p53蛋白表达。以RG7388(MDM2抑制剂)或PFT-α(p53抑制剂)与20μmol/L P22077共处理Ishikawa细胞48 h以验证USP7-MDM2-p53信号轴上下游关系。结果:USP7蛋白在子宫内膜癌组织和细胞中高表达,且Ishikawa细胞中USP7蛋白表达量最高,因此,选择Ishikawa细胞为研究对象。与NC组比较,P22077组Ishikawa细胞OD 450值、克隆形成率、S期和G 2/M期细胞数、USP7、CyclinD1、CDK2、MDM2蛋白表达降低,细胞凋亡率、G_(0)/G_(1)期细胞数、p53、Bax蛋白表达升高(P<0.05);与NC组、pcDNA组比较,pcDNA-MDM2组Ishikawa细胞OD 450值、克隆形成率、S期和G 2/M期细胞数、USP7、CyclinD1、CDK2、MDM2蛋白表达升高,细胞凋亡率、G_(0)/G_(1)期细胞数、p53、Bax蛋白表达降低(P<0.05);与P22077组、P22077+pcDNA组比较,P22077+pcDNA-MDM2组Ishikawa细胞OD 450值、克隆形成率、S期和G 2/M期细胞数、USP7、CyclinD1、CDK2、MDM2蛋白表达升高,细胞凋亡率、G_(0)/G_(1)期细胞数、p53、Bax蛋白表达降低(P<0.05)。p53为USP7-MDM2通路下游分子。结论:抑制USP7表达可能通过下调MDM2来激活p53进而抑制Ishikawa细胞增殖、促进细胞凋亡及周期停滞。Objective:To investigate the impacts of ubiquitin-specific protease 7(USP7)on the proliferation,apoptosis and cell cycle of endometrial carcinoma cells by regulating the Mdm2 p53 binding protein homolog(MDM2)-p53 axis.Methods:Western blot was applied to detect the expression of USP7 protein in human endometrial carcinoma tissue,paracancerous tissue,human endometrial epithelial cells hEEC and human endometrial carcinoma cell lines Ishikawa,HEC-1-A,KLE.Ishikawa cells were grouped into NC group,P22077(USP7 inhibitor)group,pcDNA group,pcDNA-MDM2 group,P22077+pcDNA group,and P22077+pcDNA-MDM2 group.The proliferation of Ishikawa cells was detected by CCK-8 and clonogenic assay.The apoptosis and cell cycle of Ishikawa cells were detected by flow cytometry.Western blot was applied to detect the expression of USP7,CyclinD1,cyclin dependent kinase 2(CDK2),Bcl-2 associated X protein(Bax),MDM2 and p53 protein in Ishikawa cells.Ishikawa cells were treated with RG7388(MDM2 inhibitor)or PFT-α(p53 inhibitor)and 20μmol/L P22077 for 48 h to verify the upstream and downstream relationship of USP7-MDM2-p53 signal axis.Results:USP7 protein was highly expressed in endometrial carcinoma tissues and cells,and the highest expression of USP7 protein was found in Ishikawa cells,therefore,Ishikawa cells were selected as the research object.Compared with NC group,the OD 450 value of Ishikawa cells,clone formation rate,the numbers of cells in S phase and G 2/M phase,the expression of USP7,CyclinD1,CDK2,MDM2 proteins in P22077 group were lower,and the apoptosis rate,the number of cells in G_(0)/G_(1)phase,the expression of p53 and Bax proteins were higher(P<0.05).Compared with NC group and pcDNA group,the OD 450 value of Ishikawa cells,clone formation rate,the numbers of cells in S phase and G 2/M phase,the expression of USP7,CyclinD1,CDK2,MDM2 proteins in pcDNA-MDM2 group were higher,and the apoptosis rate,the number of cells in G_(0)/G_(1)phase,the expression of p53 and Bax proteins were lower(P<0.05).Compared with P22077 group and P22077+

关 键 词：泛素特异性蛋白酶7 Mdm2 p53结合蛋白同源物(MDM2)-p53轴 子宫内膜癌 增殖 凋亡 细胞周期 

分 类 号：R737.33[医药卫生—肿瘤]