先天性小耳畸形microRNA及mRNA表达谱的变化及关键靶基因的筛选

Screening the microRNA and mRNA Expression Profile of Congenital Microtia to Choose the Target Genes

目的筛查先天性小耳畸形患者整个基因组存在表达水平异常的microRNAs(miRNAs)和mRNAs,从中找出关键靶基因,进而探讨关键靶基因表达水平的异常及其调控网络与先天性小耳畸形发病之间的关系。方法收集先天性小耳畸形患者的残耳软骨组织为研究对象,以同一患者的健侧正常耳软骨组织作为自身对照。选用Exiqon miRCURY LNATMmicroRNA Array芯片及Arraystar Human mRNA/LncRNA Microarray芯片,对实验组3例及对照组3例样本的基因组miRNAs和mRNAs表达水平进行扫描,利用差异表达的倍数变化筛选存在表达水平差异的差异miRNAs和mRNAs。通过miRanda、miRDB、miRWalk、RNA22、Targetscan这5个数据库进行检索,预测差异miRNAs调节的靶基因。将预测靶基因与差异mRNAs对应的基因进行交叉对比,寻找同时存在miRNA和mRNA表达水平差异的关键靶基因及其对应的关键miRNAs。结果本研究筛选出24个具有表达水平差异的miRNAs,同时筛选出515个表达水平差异的mRNAs;将预测到的miRNAs调节的靶基因与mRNA表达谱中的差异基因进行交叉对比,得到对应的miRNA和mRNA表达水平均存在显著性差异的关键靶基因6个(CAST、MLL、MMP13、OTUD4、PDE5A、TGOLN2)。结论初步建立了先天性小耳畸形的miRNA表达谱和mRNA表达谱,找到了先天性小耳畸形表达异常的关键靶基因;初步建立了1个与miRNA以及mRNA表达水平相关的调控网络,有利于进一步探讨关键靶基因及其调控网络与先天性小耳畸形发病的关系。

Objective To screen the microRNA and mRNA expression profile of congenital microtia to identify the differential ex- pression in mieroRNAs and mRNAs. Locking in key target genes to establish the key genes - microRNA control network and to discuss the relationship between differential expressed microRNAs and mRNAs and the etiology of congenital microtia. Methods We selected six samples from congenital microtia to study the microRNA and mRNA expression. We collected the residual ear cartilage of microtia as the research object; normal ear cartilage of patients without ear malformations as control. We Chosed Exiqon miRCURY LNATM microRNA Ar- ray and Arraystar Human mRNA/LncRNA Microarray. Fold change of differential expression were applied to screen key miRNAs and mR- NAs. miRanda,miRDB,miRWalk,RNA22 and Targetscan were used to predict target genes of key miRNAs. The predicted target genes and differential expressed genes were cross matched to find the key target genes and the corresponding key miRNAs with differential ex- pression. Results There were 24 miRNAs and 515 mRNAs with differential expression between experimental group and control group. 6 key target genes （ CAST, MLL, MMP13, OTUD4, PDE5 A, TGOLN2 ） with significant difference in mRNA and corresponding miRNA expres- sion levels were obtained by cross matching. Conclusion In this study we detected genome - wide mRNA and microRNA level of congen- ital microtia by epigenetics method, and preliminary established a key genes - microRNA control network. It is helpful to explore the fur- ther relationship between the key genes - microRNA control network and the molecular pathogenesis of congenital microtia.