人心力衰竭心肌microRNA表达谱分析及基因调控网络的初步研究

Altered gene and microRNA in heart failure patients

目的:应用人类全基因组芯片及microRNA芯片技术,获得心力衰竭(心衰)患者与正常人心肌细胞差异表达基因及microRNA,并进行功能和通路分析,构建完整的心衰基因调控网络,发现网络中起核心调控作用的基因及microRNA。方法:14例心衰患者及8例脑死亡的人正常心肌标本,分别杂交人全基因组芯片和microRNA芯片。对差异基因进行功能和通路分析,构建心衰组和正常组的基因调控网络;分析差异表达microRNA的靶基因与差异基因,构建microRNA的功能调控网络,最终得到网络中的核心基因和microRNA。结果:①与正常组比较,心衰组上调基因207条,下调150条。经KEGG通路分析,差异基因参与了显著性上调的通路20个,下调的仅有2个。构建心衰组和正常组的基因调控网络,得到22个关键基因。②与正常组比较,心衰组上调microRNA 58条,下调41条。通过搜索TargetScan数据库以及其明公司microRNA预测引擎,得到差异microRNA调控的所有靶基因22 074条。③差异microRNA靶基因与差异基因交集后得到153条基因,构建microRNA调控网络,得到7个关键microRNA(包括microRNA-200b、-181c、-340、-557、-19a、-19b及-548f),主要调节蛋白质氨基酸的磷酸化、多细胞的机体发育和免疫应答等功能。结论:心衰时心肌细胞的基因及microRNA表达谱发生了明显变化,基因调控网络存在显著不同,构建完整的心衰基因调控网络,有助于发现核心基因,深入理解心衰发生发展的分子机制,提供新的治疗靶点。

Objective：To gain altered gene and microRNA in heart failure （HF） patients by human whole geno- mewide and microRNA chip hybridization. Further, builded the global gene regulation networks, and found the nodal gene and mieroRNA followed by function and pathway analysis. Method： Human whole genomewide and microRNA chip hybridization were carried out on 14 patients with HF and 8 brain died patients without heart dis- ease, and the altered gene was selected followed function and pathway analysis to build HF and normal group gene regulation networks by analyzing altered gene and target gene of altered microRNA to find the nodal gene and mieroRNA. Result：All 357 genes and 99 microRNAs were altered （more than 2-folds） in HF and control group. Twenty pathways up-regulated and only 2 down-regulated by KEGG PATHWAY Database. Twenty-two key genes were identified by building HF and control group gene regulation networks. One hundred and fifty-three genes were identified in the intersection of target gene of altered microRNAs and altered genes, and 7 key microR- NAs （including microRNA-200b, -181c, -340, -557, -19a, -19b, -548f） were identified by building microRNA regulation network, which regulate protein amino acid phosphorylation, cellulous organism development and immu- nologic response. Conclusion： Gene expression, microRNA profile and gene regulation change markedly in HF patients. Building global gene regulation networks to find the nodal gene and microRNA is useful for understand- ing the pathogenesis of HF and providing new therapy target.