摘要
目的通过RNA测序(RNA sequencing,RNA-Seq)对先天性马蹄内翻足(congenital talipes equinovarus,CTEV)患儿血浆中游离的总RNA行全转录组测序,比较CTEV患儿与正常儿童血浆的mRNA、长链非编码RNA(long non-coding RNA,lncRNA)、环状RNA(circular RNA,circRNA)以及微小RNA(microRNA,miRNA)的差异表达,揭示CTEV的可能致病机制。方法选取3例CTEV患儿(试验组)及3例正常儿童(对照组)血浆样品,应用RNA-Seq技术分析3对血浆样品的转录组,筛选出CTEV患儿与正常儿童的差异表达基因与转录本。应用基因本体论方法对差异表达基因进行功能显著性富集分析,基于基因与基因组京都百科全书对差异基因进行显著性信号通路富集分析。结果成功构建样品的基因表达谱:试验组和对照组共有181个差异表达基因,其中58个上调基因,123个下调基因。差异表达lncRNA 23个,上调12个,下调11个。差异表达miRNA 23个,上调11个,下调12个。未筛选出差异表达circRNA。其中分泌磷酸蛋白1(secreted phosphoprotein 1,SPP1)和核糖体蛋白S27(ribosomal protein S27,RPS27)在试验组和对照组中基因表达量差异明显,差异倍数分别是82.24和0.01。结论RNA测序可对CTEV患儿和正常儿童的血浆中差异表达基因进行筛选。SPP1和RPS27可能与CTEV的发病相关。
Objective To sequence the whole transcriptomics of congenital talipes equinovarus(CTEV)by RNA sequencing(RNA-Seq)and to compare the differential expression of mRNA, long non-coding RNA(lncRNA), circular RNA(circRNA)and microRNA(miRNA)between CTEV and normal children.Methods RNA was extracted from plasma samples of 3 CTEV children(experimental group)and 3 normal children(control group). After RNA-Seq, the differentially expressed genes(DEG)and transcripts of CTEV and normal children were screened out.Gene ontology analysis and Kyoto encyclopedia of genes and genomes pathway enrichment analysis were carried out for the DEG.Results The gene expression profile was successfully constructed.There were 181 DGE in the experimental group and control group, including 58 up-regulated genes and 123 down-regulated genes.A total of 23 lncRNA, 12 up-regulated and 11 down-regulated, were differentially expressed in the present study.There were 23 miRNA differentially expressed, including 11 up-regulated and 12 down-regulated miRNA.No differential expression of circRNA was found in the present study.The expression of secreted phosphoprotein 1(SPP1)and ribosomal protein S27(RPS27)was significantly different in the experimental group and the control group with the fold change being 82.24 and 0.01, respectively.Conclusion DGE in the plasma samples of CTEV and normal children were analysed by full transcriptome sequencing.SPP1 and RPS27 may be the candidate genes of CTEV.
作者
徐晨晨
刘振江
Xu Chenchen;Liu Zhenjiang(Department of Pediatric Orthopedics,Shengjing Hospital of China Medical University,Shenyang 110004,China)
出处
《国际儿科学杂志》
2020年第6期431-435,F0003,共6页
International Journal of Pediatrics
关键词
儿童
先天性马蹄内翻足
全转录组测序
RNA测序
差异表达基因
Children
Congenital talipes equinovarus
Whole transcriptome sequencing
RNA sequencing
Differentially expressed genes
