组蛋白去乙酰化酶抑制剂CUDC-101对前列腺癌DU145细胞DNA损伤、迁移和上皮-间质转化的影响  

Effect of histone deacetylase inhibitor CUDC-101 on DNA damage,migration,and epithelial-mesenchymal transition of prostate cancer DU145 cells

作　　者：那佈其 权春姬[1] 赵芳 杨凡 肖茹 金雪梅[1] 李珍玲[1] NA Buqi;QUAN Chunji;ZHAO Fang;YANG Fan;XIAO Ru;JIN Xuemei;LI Zhenling(Department of Pathology,Affiliated Hospital,Yanbian University,Yanji 133000,China;Department of Pathology,Tiemei General Hospital of Health Industry Group,Diaobingshan 112700,China)

机构地区：[1]延边大学附属医院病理科,吉林延吉133000 [2]健康产业集团铁煤总医院病理科,辽宁调兵山112700

出　　处：《吉林大学学报（医学版）》2024年第2期400-410,共11页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金地区基金项目(81860043);国家自然科学基金青年基金项目(81902790);吉林省科技厅科技发展计划项目(20200201571JC);延边大学应用基础项目(延大科合字[2019]第27号)。

摘　　要：目的:探讨泛素化组蛋白(H2AX)在前列腺癌(PCa)和癌旁良性前列腺组织中的表达及与其PCa患者临床病理参数之间的关系,阐明新型组蛋白去乙酰化酶抑制剂(HDACi)CUDC-101对PCa的DNA损伤、迁移及上皮-间质转化(EMT)的影响。方法:肿瘤基因组谱(TCGA)数据库和UALCAN数据库检索各种癌症组织中H2AX mRNA的表达情况及其在PCa与正常前列腺组织中的表达差异,分析其表达与PCa患者临床预后的关系;采用免疫组织化学染色法检测在PCa组织和癌旁良性前列腺组织中H2AX蛋白的表达情况,分析H2AX蛋白表达与PCa患者临床病理参数的关系;体外培养DU145细胞,分为对照组、5%FBS组、5%FBS+100μmol·L^(-1)CUDC-101组和5%FBS+200μmol·L^(-1)CUDC-101组,采用细胞划痕实验和Transwell小室实验检测CUDC-101处理前后PCa DU145细胞的细胞划痕面积及迁移细胞数;免疫荧光染色法检测CUDC-101处理后PCa DU145细胞中上皮细胞标志物E钙黏蛋白(E-cadherin)和磷酸化组蛋白γ-H2AX表达情况;Western blotting法检测CUDC-101处理后EMT相关蛋白、γ-H2AX和磷酸化蛋白激酶B(p-AKT)表达情况。结果:TCGA数据库和UALCAN数据库分析,H2AX mRNA在PCa组织中高表达,并且H2AX低表达组患者的无病生存期(DFS)明显高于H2AX mRNA高表达组(P<0.001);免疫组织化学染色,在PCa组织中H2AX蛋白强阳性表达率高于在癌旁良性前列腺组织(64.34%vs 14.29%),其过表达与PCa的T分期(P=0.001)和世界卫生组织/国际泌尿科病理学会(WHO/ISUP)预后分级分组(P=0.004)有关,但与PCa患者的年龄、Gleason评分、淋巴结转移、神经和脉管浸润无关(P>0.05);免疫荧光染色法检测,与对照组和EMT诱导组比较,CUDC-101处理组E-cadherin和γ-H2AX蛋白的荧光表达增强;细胞划痕实验和Transwell小室实验检测,与对照组比较,CUDC-101处理组DU145细胞愈合面积和迁移细胞数明显下调;Western blotting法检测,与EMT诱导组比较,CUDC-101处理组E-cadherin和γ-H2Objective:To discuss the expression of ubiquitinated histone(H2AX)in the prostate cancer(PCa)and adjacent benign prostate tissues,and its relationship with the clinicopathological parameters of the PCa patients,and to clarify the effect of the novel histone deacetylase inhibitor(HDACi)CUDC-101 on the DNA damage,migration,and epithelial-mesenchymal transition(EMT)in PCa.Methods:The expression levels of H2AX mRNA in various cancer tissues were retrieved from The Cancer Genome Atlas(TCGA)and UALCAN Databases to analyze the expression differences between PCa and normal prostate tissues and its connection with the clinical prognosis of the patients with PCa;immunohistochemistry method was used to detect the expression of H2AX protein in PCa tissue and adjacent benign prostate tissue,and its relationship with the clinicopathological parameters of the PCa patients was analyzed.The DU145 cells were cultured in vitro and divided into control group,5%FBS group,5%FBS+100μmol·L^(-1) CUDC-101 group,and 5%FBS+200μmol·L^(-1) CUDC-101 group.Cell scratch assay and Transwell chamber assay were used to detect the scratch area of the cells and the number of migration cells in the PCa DU145 cells before and after treated with CUDC-101;immunofluorescence staining was used to detect the expressions of epithelial cell marker E-cadherin(E-cadherin)and phosphorylated histoneγ-H2AX in the PCa DU145 cells after treated with CUDC-101;Western blotting method was used to detect the expression levels of EMT-related protein,γ-H2AX,and phosphorylated protein kinase B(p-AKT)in the PCa DU145 cells after treated with CUDC-101.Results:The TCGA Database and UALCAN Database analysis results showed that H2AX mRNA was highly expressed in the PCa tissue,and the disease-free survival(DFS)of the patients with low expression of H2AX was longer than those patients with high expression of H2AX mRNA(P<0.001);the immunohistochemistry results showed that compared with adjacent benign prostate tissue,the rate of strong positive expression of H2AX protein in

关 键 词：CUDC-101 泛素化组蛋白 前列腺肿瘤 上皮-间质转化 DNA损伤 

分 类 号：R737.2[医药卫生—肿瘤]