mmu-miR-107调控Cacna2d1基因的mRNA水平  

Regulation of m RNA levels of Cacna2d1 by Mmu-miR-107

作　　者：阮杰[1,2,3,4] 翁亚光[5] 赵炜[1,4] 李江滨[1,2] 黄华明[1,4] 黄池荣 刘桂平[2] 刘新光[1,2,3,4] 

机构地区：[1]广东医学院衰老研究所 [2]广东医学院医学检验学院 [3]广东医学院生物化学与分子生物学研究所 [4]广东省分子诊断重点实验室,广东东莞523808 [5]重庆医科大学检验医学院,重庆400016

出　　处：《广东医学院学报》2015年第5期533-537,共5页Journal of Guangdong Medical College

基　　金：国家自然科学基金项目(No.81170327);广东省医学科研基金(No.A2014470);东莞市医疗卫生一般项目(No.201410515200159;2011108102031);广东医学院科研基金项目(No.M2013002;M2010007);广东医学院大学生创新创业训练计划项目(No.201310571009;1057113036);广东医学院建博科技创新团队项目(No.STIF201102)

摘　　要：目的生物信息学预测并实验验证mmu-miR-107的靶基因。方法利用Target Scan、miRanda、Clip-seq及miRDB预测mmu-miR-107靶基因,分别构建含有候选靶基因(Cacna2d1、Capza2、Celsr2、Csnk1g2和Tmem47)3'UTR的荧光素酶报告载体p GL3,通过双荧光素酶检测系统、时实定量PCR进一步验证预测的靶基因。结果成功构建分别含有5个候选靶基因3'UTR的荧光素酶报告载体;与mi R-NC组相比,mi R-107组共转染p GL3-Cacna2d1 3'UTR、p GL3-Cav13'UTR的荧光素酶活性均显著下降(P<0.01或0.05);其余候选靶基因Capza2、Celsr2、Csnk1g2和Tmem47的共转染组荧光素酶活性差异无统计学意义(P>0.05);过表达或下调C2C12细胞的miR-107,可有效降低或提高Cacna2d1的mRNA水平表达(P<0.05)。结论初步验证Cacna2d1是mmu-miR-107的靶基因。Objective To predict and confirm the target genes of mmu-mi R-107. Methods Target genes of mmu-mi R-107 was predicted by Target Scan, mi Randa, Clip-seq and mi RDB, the 3＇UTR of candidate target genes（Cacna2d1, Capza2, Celsr2, Csnk1g2 and Tmem47） were inserted in to the plasmid p GL3. Candidate target genes were verified by luciferase reporter assay, q-PCR. Results The luciferase reporter plasmid of the five candidate target genes were successfully constructed. Compared with mi R-NC Group, the luciferase activity of C2C12 cells co-transfected with p GL3-Cacna2d1 3＇UTR and p GL3-Cav1 3＇UTR in mi R-107 Group was decreased by 33.4%（P〈0.01 or 0.05）; there was no statistical difference in luciferase activity among the C2C12 cells co-transfected with other candidate target genes including Capza2, Celsr2, Csnk1g2 and Tmem47（P〉0.05）; Overexpressing or down-regulating mi R-107 in C2C12 cells, can effectively reduce or increase the m RNA levels of Cacna2d1（P〈0.05）. Conclusion It is preliminarily verified that Cacna2d1 might be a target gene of mmu-mi R-107.

关 键 词：mmu-miR-107 生物信息学 靶基因 Cacna2d1 

分 类 号：R34[医药卫生—基础医学]