miR-139-5p靶向Notch1抑制前列腺癌细胞上皮间质转化与转移  

miR-139-5p Targeted Notch1 to Suppress Epithelial-Mesenchymal Transition and Metastasis in Prostate Cancer Cells

作　　者：黄志扬[1] 蔡经爽[1] 周金[1] 张巍 辛军[1] 蔡雪华[2] 叶燕乐[1] HUANG Zhiyang;CAI Jingshuang;ZHOU Jin;ZHANG Wei;XIN Jun;CAI Xuehua;YE Yanle(Department of Urology,Quanzhou First Hospital Affiliated to Fujian Medical University,Quanzhou 362000,China;Department of Nursing,Quanzhou First Hospital Affiliated to Fujian Medical University,Quanzhou 362000,China)

机构地区：[1]福建医科大学附属泉州第一医院泌尿外科,泉州362000 [2]福建医科大学附属泉州第一医院护理部,泉州362000

出　　处：《福建医科大学学报》2023年第6期428-434,共7页Journal of Fujian Medical University

基　　金：福建省自然科学基金项目(2020J011279)。

摘　　要：目的 探讨miR-139-5p靶向调节Notch1表达对前列腺癌(PCa)细胞上皮间质转化(EMT)和转移能力的影响。方法 收集福建医科大学附属泉州第一医院泌尿外科手术切除的PCa组织标本,购买PCa细胞系,分别以良性前列腺增生(BPH)标本和人前列腺上皮细胞系为对照。采用qRT-PCR实验和Western-blot实验分别检测组织和细胞中miR-139-5p和Notch1的表达;采用双荧光素酶报告实验验证miR-139-5p与Notch1的靶向关系;采用Western-blot实验检测细胞EMT标志物E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)和Vimentin蛋白水平;采用Transwell实验检测细胞迁移能力;采用裸鼠尾静脉注射肿瘤细胞的方式建模,评估肺部肿瘤转移灶的情况。结果 在组织学和细胞水平上,与BPH组织比较,前列腺癌miR-139-5p低表达,而Notch1高表达,差别有统计学意义(P<0.05)。与对照组比较,高表达miR-139-5p的PC-3细胞E-cadherin表达升高,N-cadherin和Vimentin表达降低;Transwell迁移实验每视野穿膜细胞数更少;裸鼠肺转移模型中平均肺转移瘤病灶数目更少,差别有统计学意义(P<0.05)。miR-139-5p mimics转染同时过表达Notch1,与单纯miR-139-5p mimics转染比较,E-cadherin水平降低,而N-cadherin和Vimentin水平升高,Transwell迁移实验中穿膜细胞数也增加,差别有统计学意义(P<0.05)。过表达miR-139-5p降低PC-3细胞Notch1的mRNA和蛋白表达水平。双荧光素酶报告实验结果显示,野生型Notch1的PC-3细胞中,转染miR-139-5p mimics组细胞荧光活性显著低于miR-NC处理组,差别有统计学意义(P<0.05)。结论 miR-139-5p在PCa中低表达,可通过上调其表达靶向抑制Notch1,从而抑制PCa细胞EMT和远处转移。Objective To investigate the effects of miR-139-5p targeted Notch1 on epithelial-mesenchymal transition(EMT)and metastasis of prostate cancer(PCa)cells.Methods The PCa tissue samples were obtained from patients who underwent radical prostatectomy in Department of Urology,Quanzhou First Hospital Affiliated to Fujian Medical University and the PCa cell line was purchased.Samples of benign prostatic hyperplasia(BPH)and human prostatic epithelial cell line were used as the controls.qRT-PCR and Western-blot analysiswere performed to detect the expression levels of miR-139-5p and Notch1 in tissues and cells.Dual-luciferase reporterassay was performed to verify whether miR-139-5p combined with Notch1 directly.The expression levels of E-cadherin,N-cadherin and Vimentin,which were biomarkers of EMT were detected to determine the effect of miR-139-5p on EMT by Western-blot analysis.Transwell assay was used to detect the migration ability of the cells.The lung metastasis model in nude mice was established by injecting cancer cells viatail vein to determine the metastatic efficacy.Results In cancer tissue and the cellular level,the expression of miR-139-5p was lower(P<0.05),and the expression of Notch1 was higher(P<0.05)than that in the BPH group.Compared with PC-3 cells in the BPH group,the expression of E-cadherin in the group treated with miR-139-5p mimics was higher(P<0.05),while the expression levels of N-cadherin(P<0.05)and Vimentin(P<0.05)were lower.Similarly,the cells migrating through the membrane(P<0.05)and the lung metastatic lesions(P<0.05)were fewer in the group treated with miR-139-5p mimics.The expression of E-cadherin in miR-139-5p mimics+OE-Notch1 group was significantly lower(P<0.05),which the expression levels of N-cadherin(P<0.05),Vimentin(P<0.05)were higher,compared with those in miR-139-5p mimics group.Also,the cells migrating through the membrane were more in the former group(P<0.05).Overexpression of miR-139-5p decreased the mRNA(P<0.05)and protein(P<0.05)expression levels of Notch1 in PC-3 cells.The

关 键 词：miR-139-5p 前列腺癌 上皮间质转化 远处转移 

分 类 号：R737.25[医药卫生—肿瘤]