circZNF652/miR-140-3p/HMGB1通路对前列腺癌细胞增殖和迁移的影响及机制  

Effect and mechanism of circZNF652/miR-140-3p/HMGB1 pathway on cell proliferation and migration in prostate cancer

作　　者：姜华 张贺 江松松 JIANG Hua;ZHANG He;JIANG Songsong(Department of Urologic Surgery,Fifth Affiliated Hospital(Zhuhai)of Zunyi Medical University/Zhuhai Municipal Sixth People’s Hospital,Zhuhai,Guangdong 519100,China)

机构地区：[1]遵义医科大学第五附属(珠海)医院/珠海市第六人民医院泌尿外科,广东珠海519100

出　　处：《重庆医学》2025年第2期303-312,318,共11页Chongqing Medical Journal

基　　金：广东省基础与应用基础研究基金项目(2022A1515220218);贵州省科技计划项目(黔科合基础-ZK[2022]一般633);贵州省科技支撑计划(一般项目)(黔科合支撑[2023]一般262);广东省医学科研基金项目(B2023140);遵义医科大学博士科研启动基金项目(BS2022-01)。

摘　　要：目的 探讨环状RNA(circRNA)circZNF652在前列腺癌(PCa)组织及PCa细胞系中的表达及其对增殖和迁移的影响和可能机制。方法 通过基因芯片技术检测在PCa和良性前列腺增生(BPH)患者血浆组织中差异表达的circRNA。实时荧光定量PCR(qPCR)测定circZNF652在PCa细胞系(22RV1、LNCaP、DU145、PC3)和正常前列腺上皮细胞(RWPE-1)中的表达情况。分析circZNF652不同表达水平与PCa患者总生存(OS)时间及临床病理特征的相关性。细胞计数试剂盒-8(CCK-8)实验、克隆形成实验、Transwell实验、细胞划痕实验、EdU细胞增殖实验分析干扰circZNF652对PCa细胞增殖、克隆形成、侵袭和迁移能力的影响。在线数据库TargetScan预测circZNF652和miR-140-3p之间的结合位点,双荧光素酶报告基因实验,RNA pull-down实验验证circZNF652和miR-140-3p的相互作用,并通过在PC3和DU145细胞中干扰circZNF652,以及在PCa和BPH患者血浆中进一步验证;starBase2.0预测miR-140-3p和高迁移率族蛋白B1(HMGB1)之间的结合位点;在PC3和DU145细胞中检测过表达miR-140-3p后HMGB1表达水平变化。Western blot检测PCa、BPH患者血浆,PC3、DU145、RWPE-1细胞中HMGB1表达水平,双荧光素酶报告基因实验、RNA免疫共沉淀(RIP)和RNA pull-down实验验证miR-140-3p和HMGB1的相互作用;并检测转染不同构建物(si-NC、si-circZNF652#1、si-circZNF652#1+inhibitor NC、si-circZNF652#1+miR-140-3p inhibitor)后的PC3和DU145细胞中HMGB1的表达,以进一步证实miR-140-3p直接调控HMGB1。结果 基因芯片技术及qPCR结果表明,PCa患者血浆中circZNF652的表达水平较BPH患者升高,与低级别PCa和BPH患者比较,高级别PCa患者血浆中circZNF652表达水平上调(P<0.001、P<0.01)。和RWPE-1细胞比较,22RV1、LNCaP、DU145、PC3细胞中circZNF652表达水平上调(P<0.05、P<0.05、P<0.001、P<0.01);circZNF652高表达的PCa患者OS时间较circZNF652低表达的PCa患者更短。临床病理特征分析显示,circZNF652表达与PCObjective To investigate the expression of circZNF652 in prostatic cancer(PCa)tissues and PCa cell lines,and its impact and possible mechanism on proliferation and migration.Methods Differential expression of circRNAs in plasma tissues from PCa patients and benign prostatic hyperplasia(BPH)patients were detected by using gene chip technology.The expression levels of circZNF652 in PCa cell lines(22RV1,LNCaP,DU145,PC3)and normal prostate epithelial cells were measured by quantitative real-time PCR(qPCR).The correlation between different circZNF652 expression levels with the overall survival(OS)time and clinicopathological features of PCa patient were analyzed.The effects of interfering circZNF652 on PCa cell proliferation,colony formation,invasion and migration ability were assessed through CCK-8 assay,colony formation assay,Transwell assay,wound healing assay and EdU cell proliferation assay.The online database TargetScan was used to predict the binding sites between circZNF652 and miR-140-3p,and the dual-luciferase reporter gene assay and RNA pull-down assay confirmed the interaction of circZNF652 and miR-140-3p.This interaction was further validated in PC3 and DU145 cells by interfering circZNF652 as well as in blood samples from PCa and BPH patients.StarBase predicted the binding sites between miR-140-3p and HMGB1,and the HMGB1 expression levels were tested after miR-140-3p overexpression in PC3 and DU145 cells.Western blot was used to detect the HMGB1 expression levels in PCa and BPH patient,PCa cell lines(DU145,PC3)as well as RWPE-1 cells.The dual-luciferase reporter gene assay,RNA immunoprecipitation(RIP)and RNA pull-down were conducted to verified the interaction between miR-140-3p and HMGB1,and the HMGB1expression in PC3 and DU145 cells were detected after transfecting different constructs(si-NC,si-circZNF652#1,si-circZNF652#1+inhibitor NC,si-circZNF652#1+miR-140-3p inhibitor)to further confirming that miR-140-3p directly regulates HMGB1.Results The gene chip and qRT-PCR results showed that the expression

关 键 词：circZNF652 miR-140-3p 高迁移率族蛋白B1 前列腺癌 

分 类 号：R737.25[医药卫生—肿瘤]