肝癌细胞miR-326介导的ACRBP转录抑制的研究  

Study on miR-326-mediated ACRBP transcriptional inhibition in hepatoma carcinoma cell

作　　者：包晴 李冰莹 刘俊棋 柳永清 梁晓乐[3] 谢小薰[1] 葛盈盈[1] Bao Qing;Li Bingying;Liu Junqi;Liu Yongqing;Liang Xiaole;Xie Xiaoxun;Ge Yingying(Guangxi Medical University Human Anatomy,Histology and Embryology,School of basic Medical Sciences,Nanning 530022,China;Guangxi Medical University Clinical medicine,School of basic Medical Sciences,Nanning 530022,China;Guangxi Medical University Laboratory of Basic Medicine,School of basic Medical Sciences,Nanning 530022,China)

机构地区：[1]广西医科大学基础医学院人体解剖与组织胚胎学,南宁530022 [2]广西医科大学基础医学院临床医学,南宁530022 [3]广西医科大学基础医学院基础医学中心实验室,南宁530022

出　　处：《广西医科大学学报》2020年第4期605-611,共7页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.81960453);广西自然科学基金资助项目(No.2018GXNSFBA28118);广西自然科学基金资助项目(No.2018GXNSFAA050151)。

摘　　要：目的:探究miR-326在人肝癌细胞株HepG2中对ACRBP表达的潜在调控作用。方法:运用TargetScan、miRanda和miRDB三个在线基因预测工具预测miR-326与ACRBP 3’UTR之间的保守结合位点;双荧光素酶报告系统分析ACRBP3’UTR保守结合位点突变后对miR-326与ACRBP结合能力的影响;通过慢病毒转染的方法获得稳定高表达miR-326的HepG2肝癌细胞株;利用实时荧光定量聚合酶链反应(qPCR)和Western blotting检测miR-326上调对ACRBP mRNA和蛋白表达的影响。结果:在人肝癌细胞株HepG2中,miR-326可结合于ACRBP mRNA 3’UTR的保守位点,并抑制该基因mRNA翻译,从而抑制ACRBP的表达。结论:miR-326在HepG2中负向调控ACRBP的表达。Objective: To explore the potential regulatory effect of miR-326 on ACRBP expression in HCC cell line--HepG2.Methods: Three online gene prediction tools--TargetScan,miRanda,and miRDB--were used to predict the conservative binding site between miR-326 and ACRBP 3’UTR.Dual-luciferase reporter system was used to analysis the effect of ACRBP 3’UTR conservative binding site mutation on the binding ability of miR-326 to ACRBP.HepG2 hepatoma cell line stably and highly expressing miR-326 was obtained by lentivirus transfection.The effect of up-regulation of miR-326 on ACRBP mRNA and protein expression were detected by real-time fluorescence quantitative polymerase chain reaction(qPCR)and Western blotting.Results: In human hepatoma cell line HepG2,miR-326 could bind to the conservative site of ACRBP mRNA 3’UTR and inhibited the mRNA translation of the gene,thereby inhibited the expression of ACRBP.Conclusion: miR-326 negatively regulates the expression of ACRBP in HepG2.

关 键 词：miR-326 ACRBP 肝癌 

分 类 号：R321[医药卫生—人体解剖和组织胚胎学]