Chen Chong, Tan Haining, Bi Jiaqi, Li Zheng, Rong Tianhua, Lin Youxi, Sun Liang, Li Xingye, Shen Jianxiong
Department of Orthopedic Surgery, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing, China.
Beijing Zhongke Jingyun Technology Company Ltd., Beijing, China.
Cell Physiol Biochem. 2018;48(5):2134-2146. doi: 10.1159/000492556. Epub 2018 Aug 15.
BACKGROUND/AIMS: Congenital scoliosis (CS) is a result of anomalous development of vertebrae and is frequently associated with somitogenesis malformation. Although noncoding RNAs (ncRNAs) have been recently determined to be involved in the pathogenesis of CS, the competing endogenous RNA (ceRNA) regulatory networks in CS remain largely unknown.
Sequencing was conducted to explore the ncRNA expression profiles in rat embryos (gestation day 9) following vitamin A deficiency (VAD) (n = 9 for the vitamin A deficiency-induced congenital scoliosis (VAD-CS) group and n = 4 for the control group). Real-time reverse transcriptase polymerase chain reaction (RT-PCR) was conducted to verify the expression levels of selected mRNAs, long noncoding RNAs (lncRNAs), circular RNAs (circRNAs), and microRNAs (miRNAs). Bioinformatics analysis was used to discover the possible relationships and functions of the ceRNAs.
A total of 749 mRNAs, 56 miRNAs, 685 lncRNAs, and 70 circRNAs were identified to have significantly different expression levels in the two groups. Wnt, PI3K-ATK, FoxO, EGFR, and mTOR were found to be the most significant pathways involved in VAD-CS pathogenesis. The circRNA/miRNA/mRNA and lncRNA/miRNA/mRNA networks of CS were built, and the gene expression mechanisms regulated by ncRNAs were unveiled via the ceRNA regulatory networks.
We comprehensively identified ceRNA regulatory networks of embryonic somite development in VAD-CS as well as revealed the contribution of different ncRNA expression profiles. Our data demonstrate the association between mRNAs and ncRNAs in the pathogenic mechanism of CS.
背景/目的：先天性脊柱侧凸（CS）是椎体异常发育的结果，常与体节发生畸形相关。尽管最近已确定非编码RNA（ncRNA）参与CS的发病机制，但CS中的竞争性内源性RNA（ceRNA）调控网络仍 largely未知。
进行测序以探索维生素A缺乏（VAD）后大鼠胚胎（妊娠第9天）中的ncRNA表达谱（维生素A缺乏诱导的先天性脊柱侧凸（VAD-CS）组n = 9，对照组n = 4）。进行实时逆转录聚合酶链反应（RT-PCR）以验证所选mRNA、长链非编码RNA（lncRNA）、环状RNA（circRNA）和微小RNA（miRNA）的表达水平。生物信息学分析用于发现ceRNA的可能关系和功能。
共鉴定出749种mRNA、56种miRNA、685种lncRNA和70种circRNA在两组中具有显著不同的表达水平。发现Wnt、PI3K-ATK、FoxO、EGFR和mTOR是参与VAD-CS发病机制的最显著途径。构建了CS的circRNA/miRNA/mRNA和lncRNA/miRNA/mRNA网络，并通过ceRNA调控网络揭示了ncRNA调控的基因表达机制。
我们全面鉴定了VAD-CS中胚胎体节发育的ceRNA调控网络，并揭示了不同ncRNA表达谱的作用。我们的数据证明了mRNA与ncRNA在CS致病机制中的关联。
