miR-194-5p靶向调控BCKDHA基因的表达  

miR-194-5p Targets BCKDHA Gene to Regulate Its Expression

作　　者：张莹姣 乔利英[1] 景炅婕[1] 潘洋洋[1] 马元[1] 郭云利 魏琼 刘文忠[1] 

机构地区：[1]山西农业大学动物科技学院,太谷030801 [2]左云县畜牧兽医服务中心,左云037100

出　　处：《畜牧兽医学报》2017年第8期1416-1423,共8页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金项目(31372292)

摘　　要：旨在预测并验证调控支链氨基酸分解代谢的限速酶支链α-酮酸脱氢酶复合体(Branched-chainα-ketoacid dehydrogenase complex,BCKDH)中α亚基即BCKDHA基因表达的关键miRNA。本研究利用3个在线软件预测与该基因具有靶标关系的miRNAs。利用荧光定量PCR检测小鼠3T3-L1前脂肪细胞分化过程中和超表达后关键miRNA与BCKDHA的表达量。结果表明,miR-194-5p是调控BCKDHA表达的一个关键miRNA,其种子序列与BCKDHA3′UTR第190~196位碱基完全互补。MiR-194-5p的表达随分化时间呈下降趋势,而BCKDHA mRNA则呈上升趋势。双荧光素酶报告结果也显示二者具有靶标关系,miR-194-5p下调BCKDHA的表达。超表达miR-194-5p后,BCKDHA的表达显著下调(P<0.05)。由此证明,miR-194-5p通过与BCKDHA基因3′UTR结合抑制其表达。This study aimed to predict and validate the crucial miRNA regulating BCKDHA gene expression, where BCKDHA was α subunit of BCKDH （Branched-chain α-ketoacid dehydrogenase complex）, which was a key rate-limiting enzyme in the catabolism of branched-chain amino acids. Three online softwares were used to predict miRNAs targeting BCKDHA. QPCR was used to detect expressions of the crucial miRNA and BCKDHA mRNA in 3T3-L1 cells during its differentiation and after over-expressing the crucial miRNA, respectively. The results showed that miR-194-5p was a key miRNA regulating BCKDHA expression with fully complementation between its seed sequence and the 190-196th bases of BCKDHA 3＇UTR. The expression of miR-194-5p decreased while that of BCKDHA mRNA increased with differentiation. The target relationship was also validated by dual luciferase reporter results which showed that miR-194-5p negatively regulated the expression of BCKDHA gene. The expression of BCKDHA mRNA was significantly reduced（P〈0.05） after over-expressing miR-194-5p. It is concluded that miR-194-5p inhibits the expression of BCKDHA by targeting its 3＇ UTR.

关 键 词：miR-194-5p BCKDHA基因 miRNA预测 3T3-L1 靶基因验证 

分 类 号：S813.1[农业科学—畜牧学]