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.
J Cell Mol Med. 2019 Jul;23(7):4582-4591. doi: 10.1111/jcmm.14355. Epub 2019 May 2.
Congenital scoliosis (CS) is the result of anomalous vertebrae development, but the pathogenesis of CS remains unclear. Long non-coding RNAs (lncRNAs) have been implicated in embryo development, but their role in CS remains unknown. In this study, we investigated the role and mechanisms of a specific lncRNA, SULT1C2A, in somitogenesis in a rat model of vitamin A deficiency (VAD)-induced CS. Bioinformatics analysis and quantitative real-time PCR (qRT-PCR) indicated that SULT1C2A expression was down-regulated in VAD group, accompanied by increased expression of rno-miR-466c-5p but decreased expression of Foxo4 and somitogenesis-related genes such as Pax1, Nkx3-2 and Sox9 on gestational day (GD) 9. Luciferase reporter and small interfering RNA (siRNA) assays showed that SULT1C2A functioned as a competing endogenous RNA to inhibit rno-miR-466c-5p expression by direct binding, and rno-miR-466c-5p inhibited Foxo4 expression by binding to its 3' untranslated region (UTR). The spatiotemporal expression of SULT1C2A, rno-miR-466c-5p and Foxo4 axis was dynamically altered on GDs 3, 8, 11, 15 and 21 as detected by qRT-PCR and northern blot analyses, with parallel changes in Protein kinase B (AKT) phosphorylation and PI3K expression. Taken together, our findings indicate that SULT1C2A enhanced Foxo4 expression by negatively modulating rno-miR-466c-5p expression via the PI3K-ATK signalling pathway in the rat model of VAD-CS. Thus, SULT1C2A may be a potential target for treating CS.
先天性脊柱侧凸(CS)是异常椎体发育的结果,但 CS 的发病机制尚不清楚。长链非编码 RNA(lncRNA)已被涉及到胚胎发育,但它们在 CS 中的作用尚不清楚。在这项研究中,我们研究了一个特定的 lncRNA,SULT1C2A,在维生素 A 缺乏(VAD)诱导的 CS 大鼠模型中的体节发生中的作用和机制。生物信息学分析和定量实时 PCR(qRT-PCR)表明,SULT1C2A 的表达在 VAD 组中下调,同时 rno-miR-466c-5p 的表达增加,但 Foxo4 和体节发生相关基因(如 Pax1、Nkx3-2 和 Sox9)的表达减少在妊娠第 9 天(GD)。荧光素酶报告和小干扰 RNA(siRNA)测定表明,SULT1C2A 作为竞争性内源性 RNA 通过直接结合抑制 rno-miR-466c-5p 的表达,rno-miR-466c-5p 通过结合其 3'非翻译区(UTR)抑制 Foxo4 的表达。通过 qRT-PCR 和 northern blot 分析检测到 SULT1C2A、rno-miR-466c-5p 和 Foxo4 轴在 GD 3、8、11、15 和 21 时的时空表达发生动态变化,同时 AKT 磷酸化和 PI3K 表达也发生变化。综上所述,我们的研究结果表明,SULT1C2A 通过负调控 rno-miR-466c-5p 的表达增强 Foxo4 的表达,从而在 VAD-CS 大鼠模型中通过 PI3K-AKT 信号通路。因此,SULT1C2A 可能是治疗 CS 的潜在靶点。