let-7b介导IGF2BP3表达抑制鸡成肌细胞增殖效果的研究

摘要

为探讨miRNA let-7b介导IGF2BP3对鸡成肌细胞增殖的调控作用,利用let-7b过表达载体转染成肌细胞,通过双荧光素酶靶标验证、MTT、qPCR、western blotting和ELISA分析,测定了相关基因的表达情况.结果表明:(1)采用双荧光素酶报告系统和突变实验证明let-7b作用于IGF2BP3的3'UTR.(2)let-7b在成肌细胞中过表达后,细胞增殖受到一定程度的影响,与空质粒组相比,细胞数量呈现下降的趋势.(3) let-7b可使靶基因IGF2BP3的mRNA表达量下调2.33倍,差异显著(P＜0.05),对IGF2、IGF1的mRNA表达量的影响则差异不显著(P＞0.05).研究表明,miRNA let-7b主要是通过靶向调控IGF2BP3的mRNA和蛋白质表达水平,并进一步影响IGF2的运输、稳定和迁移功能,从而影响鸡成肌细胞的增殖.%In order to investigate the role of miRNA let-7b mediated by IGF2BP3 in the proliferation of chicken myoblasts,the let-7b overexpression vector was transfected into myoblasts and verified by MTT,qPCR,western blotting and ELISA,the expression of related genes was determined.The results showed that:(1) The double-luciferase reporter system and the mutant experiments demonstrate that let-7b acts on the 3'UTR of IGF2BP3.(2) Let-7b overexpression in myoblasts,cell proliferation was affected to a certain extent,compared with the empty plasmid group,the number of cells showed a downward trend.(3) Let-7b reduced the mRNA expression of target gene IGF2BP3 by 2.33 times,and the difference was significant (P＜0.05),and there was no significant difference in the mRNA expression of IGF2 and IGF1 (P＞0.05).(4) Comprehensive analysis,miRNA let-7b is mainly through the regulation of IGF2BP3 mRNA and protein expression levels,and further affect the transport,stability and migration of IGF2 function,thus affecting the proliferation of chicken myoblasts.