miR-146a-5p靶向SMAD4抑制人前列腺癌细胞系PC-3增殖和侵袭  

miR-146a-5p inhibits proliferation and invasion of prostate cancer cell line PC-3 by targeting SMAD4

作　　者：刘彼得 李循[1] 王书恒 靳宏勇[1] 张小安[1] 李九智[1] LIU Bide;LI Xun;WANG Shuheng;JIN Hongyong;ZHANG Xiao an;LI Jiuzhi(Department of Urology,Laboratory of Urology,People's Hospital of Xinjiang Uygur Autonomous Region,Urumqi 830001,China)

机构地区：[1]新疆维吾尔自治区人民医院泌尿中心泌尿外科研究室,新疆乌鲁木齐830001

出　　处：《基础医学与临床》2023年第8期1215-1221,共7页Basic and Clinical Medicine

基　　金：新疆维吾尔自治区科学技术厅自治区自然科学基金(2017D01C102)。

摘　　要：目的探究miR-146a-5p可否靶向调控SMAD4对前列腺癌细胞增殖、侵袭的影响及其机制。方法RT-qPCR检测前列腺癌组织及各细胞系中miR-146a-5p的表达量,分析其表达与Gleason评分的关系;MTT实验、BrdU实验、集落生成实验、划痕实验、Transwell小室法及裸鼠成瘤实验分析miR-146a-5p对前列腺癌细胞增殖、成瘤、迁移及侵袭等能力的影响;RT-qPCR检测组织SMAD4的表达量,分析其与miR-146a-5p表达量的关系;荧光素酶报告基因实验分析miR-146a-5p与SMAD4的靶向关系,功能回复实验验证miR-146a-5p/SMAD4信号轴在前列腺癌中的作用;Western blot检测miR-146a-5p及SMAD4对细胞核内SMAD2/SMAD3复合体表达量的影响,并通过荧光素酶报告基因实验及染色质免疫共沉淀实验探究miR-146a-5p/SMAD4/SMAD2/SMAD3信号轴对TIM3的靶向调控作用。结果miR-146a-5p在前列腺癌组织及细胞系中低表达(P<0.05),其表达量与Gleason评分负相关(P<0.05),且在PC-3细胞中的表达量最低;miR-146a-5p抑制PC-3细胞的增殖及侵袭能力(P<0.05);SMAD4为miR-146a-5p的下游靶基因;SMAD4促进SMAD2/SMAD3复合体入核并靶向激活TIM3。结论miR-146a-5p靶向调控SMAD4抑制PC-3细胞的增殖及侵袭,SMAD2/SMAD3/TIM3信号通路可能为其下游机制。Objective To explore the inhibition effect and mechanism of miR-146a-5p on proliferation and invasion of prostate cancer(PCa)cell line PC-3 by targeting SMAD4.Methods RT-qPCR was used to detect the expression of miR-146a-5p in PCa tissues and cell lines.The relevance of miR-146a-5p expression with Gleason score was also analyzed.MTT,BrdU experiment,cell colony formation experiment,scratch experiment,Transwell assay and nude mouse xenograft model experiment were conducted to detect the effect of miR-146a-5p on cell proliferation,tumorigenicity,migration and invasion.The expression of SMAD4 in PCa tissues was detected by RT-qPCR,and the targeting relationship of SMAD4 and miR-146a-5p was confirmed by double luciferase reporter gene assay and rescue experiment.Western blot was used to detect the expression of SMAD2/SMAD3 complex in nucleus affected by miR-146a-5p and SMAD4.Finally,double luciferase reporter gene assay and ChIP experiment were performed to examine the targeting regulation of TIM3 by miR-146a-5p/SMAD4/SMAD2/SMAD3 signaling axis.Results miR-146a-5p was low expressed in PCa tissues and cell lines;its expression was negatively correlated to Gleason score and had the lowest expression in PC-3 cells.miR-146a-5p inhibited the proliferation and invasion of PC-3 cells by targeting SMAD4.SMAD2/SMAD3/TIM3 axis seemed to be the downstream mechanism of miR-146a-5p/SMAD4 signaling pathway.Conclusions miR-146a-5p can inhibit the proliferation and invasion of PC-3 cells by targeting SMAD4,and the downstream mechanism might be related to the SMAD2/SMAD3/TIM3 signaling pathway.

关 键 词：前列腺癌 miR-146a-5p SMAD4 TIM3 

分 类 号：R737.25[医药卫生—肿瘤]