摘要
目的 AKT信号通路在心力衰竭的病理机制中扮演重要角色,本研究旨在筛选调节AKT信号通路的microRNA,为进一步研究microRNA在心力衰竭发生发展中的作用奠定基础。方法首先用Gene-SetEnrichmentAnalysis方法及Targetscan和RNAhybrid软件寻找可能调节AKT信号通路的microRNA,然后用双荧光素酶报告系统验证microRNA对AKT信号通路基因3'-UTR的调节作用。结果 GeneSetEnrich-mentAnalysis分析提示:miR-15/16、miR-21、miR-29、miR-103、miR-126、miR-128、miR-129、miR-200和miR-493共9个microRNA家族可能调节AKT信号通路;利用Targetscan和RNAhybrid软件反向验证以上9个候选microRNAs,miR-29与AKT信号通路中的10个基因(TGFβ3、HDGF、VEGF、LEP、PDGFRB、PIK3RI、AKT1、AKT2、AKT3和HIF1A)的3'-UTR杂交分值较高,进一步实验验证发现TGFβ3、VEGF、LEP、AKT1、AKT2、AKT3和HIF1A都是miR-29的靶基因。结论 AKT信号通路中的多个基因都是miR-29的靶基因,可受到miR-29的负调节作用。
Objective AKT signal pathway plays an important role in the pathogenesis of heart failure.This study intends to screen the microRNAs that participate in the regulation of AKT signal pathway,and lays a foundation for the investigation of microRNAs implicated in the development of heart failure.Methods First,the candidate microRNAs possibly involved in the regulation of AKT signal pathway was predicted by using GeneSet Enrichment Analysis(GEA),TargetScan,and RNAhybrid.Then,we verified the regulation of candidate microRNAs on the candidate genes was verified using a Dual Luciferase Reporter System(DLRS).Results GEA showed that miR-15/16,miR-21,miR-29,miR-103,miR-126,miR-128,miR-129,miR-200 and miR-493 may be implicated in the regulation of AKT pathway.Further analysis of the match degree of the 9 microRNAs with the 10 genes in the AKT pathway using TargetScan and RNAhybrid software indicated that TGFβ3,HDGF,VEGF,LEP,PDGFRB,PIK3RI,AKT1,AKT2,AKT3 and HIF1A may be the target genes of miR-29.Experimental verification with DLRS suggested that TGFβ3,VEGF,LEP,AKT1,AKT2,AKT3 and HIF1A are the target genes of miR-29.Conclusions Several genes in the AKT signal pathway are the targets of miR-29 and may be down-regulated by miR-29.
出处
《中华临床医师杂志（电子版）》
CAS
2012年第14期3871-3874,共4页
Chinese Journal of Clinicians(Electronic Edition)
基金
国家自然科学基金(81100244/H2501)
广东省优秀博士论文作者资助(sybzzxm201127)
高等学校博士学科点专项科研基金(20104423120002)
