微小RNA-1246靶向糖原合成酶激酶3β激活Wnt/β-连环蛋白信号通路促进前列腺癌细胞的迁移、侵袭和增殖  

MicroRNA-1246 targeting glycogen synthase kinase-3β promotes prostate cancer migration, invasion, and proliferation by activating Wnt/β-catenin signaling pathway

作　　者：齐强元 刘皎林 陈西良 平浩 Qi Qiangyuan;Liu Jiaolin;Chen Xiliang;Ping Hao(Department of Urology,Linyi City Central Hospital,Linyi 276400,China;Department of Urology,Beijing Tongren Hospital Affiliated to Capital Medical University,Beijing 100730,China)

机构地区：[1]临沂市中心医院泌尿外科,临沂276400 [2]首都医科大学附属北京同仁医院泌尿外科,北京100730

出　　处：《中华实验外科杂志》2024年第1期83-86,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目(82072833)。

摘　　要：目的探讨微小RNA(miR)-1246靶向糖原合成酶激酶3β(GSK3β)对前列腺癌(PCa)细胞迁移、侵袭和增殖的作用及分子机制。方法采用miRNA-Seq技术检测PCa组织及癌旁组织中差异表达的miRNA;采用实时荧光定量聚合酶连反应(qRT-PCR)技术检测PCa患者癌组织和血清中miR-1246的表达水平;采用生物信息学分析、荧光素酶报告基因实验、qRT-PCR、蛋白质印迹法分析miR-1246对GSK3β的靶向调控关系。人PCa细胞DU145和LNCaP分别转染miR-1246类似物(mimics)/阴性对照(NC mimics)或miR-1246抑制剂(inhibitor)/阴性对照(NC inhibitor),通过细胞计数试剂盒(CCK-8)法检测细胞的增殖能力,通过划痕实验和Transwell迁移实验检测细胞的迁移侵袭能力。两组间比较采用独立样本t检验。结果miRNA-Seq结果显示,与癌旁组织比较,PCa组织中有15个miRNA表达显著升高,51个miRNA表达显著降低,其中miR-1246在PCa患者组织及血清中表达显著升高(组织:7.93±2.39比1.00±0.16,t=10.070,P<0.01;血清:15.23±2.77比1.01±0.09,t=17.770,P<0.01)。生物信息分析结果显示,miR-1246与GSK3β有互补的核苷酸序列,miR-1246 mimics+WT-GSK3β组细胞荧光素酶活性显著低于NC mimics+WT-GSK3β组(0.58±0.09比1.00±0.14,t=8.128,P<0.01)。蛋白质免疫印迹结果显示,miR-1246 inhibitor组GSK3β的表达明显高于NC inhibitor组(2.43±0.29比1.00±0.09,t=10.180,P<0.01),miR-1246 mimics组GSK3β的表达明显低于NC mimics组(0.51±0.07比1.01±0.12,t=5.120,P<0.05)。在DU145和LNCaP细胞中,miR-1246 mimics组细胞增殖能力、细胞迁移率和细胞侵袭数显著高于NC mimics组,miR-1246 inhibitor组细胞增殖能力、细胞迁移率和细胞侵袭数显著低于NC inhibitor组。蛋白质免疫印迹实验结果显示,miR-1246 inhibitor组Wnt3a和β-catenin的表达低于NC inhibitor组(Wnt3a:0.39±0.05比1.00±0.12,t=6.620,P<0.01;β-catenin:0.54±0.09比0.98±0.10,t=6.032,P<0.01)。结论miR-1246通过抑制GSK3β,激活Wnt/β-catenin信号通路,促�Objective To investigate the effect and molecular mechanism of microRNA(miR)-1246 on the migration,invasion and proliferation of prostate cancer(PCa)cells by targeting glycogen synthase kinase 3β(GSK3β).Methods MiRNA-Seq was used to detect the differentially expressed miRNAs in PCa tissues and adjacent tissues.The expression levels of miR-1246 in cancer tissues and serum of PCa patients were detected by real-time fluorescence quantitative polymerase chain reaction(qRT-PCR).Bioinformatics analysis,luciferase reporter gene experiment,qRT-PCR and Western blotting were used to analyze the targeted regulation relationship between miR-1246 and GSK3β.Human PCa cells DU145 and LNCaP were transfected with miR-1246 mimics/negative control(NC mimics)or miR-1246 inhibitor/negative control(NC inhibitor),respectively.The proliferation ability of cells was detected by cell counting kit-8(CCK-8)method,and the migration and invasion ability of cells were detected by scratch test and Transwell migration test.Independent sample t test was used for comparison between the two groups.Results The results of miRNA-Seq showed that 15 miRNAs were significantly up-regulated and 51 miRNAs were significantly down-regulated in PCa tissues compared with adjacent tissues.Among them,miR-1246 was significantly up-regulated in PCa tissues and serum(tissue:7.93±2.39 vs.1.00±0.16,t=10.070,P<0.01;Serum:15.23±2.77 vs.1.01±0.09,t=17.770,P<0.01).Bioinformatics analysis showed that miR-1246 and GSK3βhad complementary nucleotide sequences.The luciferase activity of miR-1246 mimics+WT-GSK3βgroup was significantly lower than that of NC mimics+WT-GSK3βgroup(0.58±0.09 vs.1.00±0.14,t=8.128,P<0.05).The results of Western blotting showed that the expression of GSK3βin miR-1246 inhibitor group was significantly higher than that in NC inhibitor group(2.43±0.29 vs.1.00±0.09,t=10.180,P<0.01),and the GSK3βexpression in miR-1246 mimics group was significantly lower than that in NC mimics group(0.51±0.07 vs.1.01±0.12,t=5.120,P<0.05).In DU145 and LNCaP

关 键 词：前列腺癌 糖原合成酶激酶3Β 迁移 侵袭 增殖 

分 类 号：R737.25[医药卫生—肿瘤]