MicroRNA-30a在胃癌组织的表达分析及其靶基因成纤维活化蛋白α的鉴定  

Identification of fibroblast activation protein α as target of microRNA-30a in gastric cancer

作　　者：龚莉[1] 左钱飞[1] 余婷[1] 汪莉娜[1] 肖斌[1] 曾浩[1] 

机构地区：[1]第三军医大学药学系微生物与生化药学教研室暨国家免疫生物制品工程技术研究中心,重庆400038

出　　处：《第三军医大学学报》2016年第6期560-563,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金项目(81202310);重庆市青年科技人才培养项目(CSTC2014kjrc-qnrc10008)~~

摘　　要：目的分析microRNA-30a(miR-30a)在胃癌中的表达水平并鉴定其新的靶基因。方法选取29例胃癌患者的肿瘤组织和癌旁组织,利用qRT-PCR检测miR-30a的表达。通过miRNA靶基因预测数据库Target Scan预测miR-30a的靶基因。构建含有miR-30a结合位点的3'UTR片段的荧光素酶载体(p MIR-FAPα),通过荧光素酶报告实验对预测靶基因进行检测。利用Western blot和qRT-PCR对预测靶基因成纤维活化蛋白α(fibroblast activation proteinα,FAPα)的表达进行检测。结果 miR-30a在胃癌组织样本中的表达明显下调,通过生物信息学预测(Target Scan)和荧光素酶实验表明miR-30a mimic可与FAPα基因结合,qRT-PCR和Western blot结果表明miR-30a mimic可抑制FAPαmRNA和蛋白质水平的表达。结论 miR-30a在人胃癌组织样本中表达下调,FAPα是其靶基因。Objective To investigate the expression of microRNA-30a （miR-30a） in human gastric cancer （GC） tissues and identify its direct target gene. Methods The expression of miR-30a in the tumor tissues and paratumor tissues of 29 patients with GC was detected by qRT-PCR. The potential target genes of miR-30a were predicted with TargerScan database. Luciferase reporter plasmid pMIR-fibroblast activation protein α （FAPα） with FAPα 3′UTR as the binding site of miR-30a was constructed. The predicted target gene （FAPα） of miR-30a was identified by dual luciferase activation assay. The mRNA and protein expression levels of FAPα were assessed by qRT-PCR and Western blotting, respectively. Results The expression of miR-30a was significantly decreased in the tumor tissues compared with the paratumor tissues. Low expression of miR-30a in the tumor tissues was significantly correlated with the clinical stage. MiR-30a mimic significantly inhibited luciferase activation （P〈0.05） in the HEK293 cells, which proved that miR-30a could target FAPα 3′UTR. Western blotting and qRT-PCR showed that miR-30a inhibited the mRNA and protein expression levels of FAPα. Conclusion MiR-30a is down-regulated in GC, and FAPα is its target gene.

关 键 词：胃癌 miR-30a 成纤维活化蛋白α 

分 类 号：R394.3[医药卫生—医学遗传学] R730.23[医药卫生—基础医学]