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Abstract：
Ankylosis spondylitis (AS) is a disease mainly characterized by sacroiliac joint and spinal attachment point inflammation. Long non-coding RNA (lncRNA) plays a key role in the progression of many diseases. However, few studies have been conducted on the function of lncRNA maternally expressed gene 3 (MEG3) in AS. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to measure the relative levels of MEG3, microRNA let-7i, sclerostin (SOST), and inflammatory cytokines. Dual-luciferase reporter assay, RNA immunoprecipitation (RIP) assay and biotin-labeled RNA pull-down assay were used to confirm the interaction between MEG3 and let-7i or let-7i and SOST. In addition, western blot (WB) analysis was performed to detect the protein levels of osteogenesis markers and SOST. The expression levels of MEG3 and SOST were decreased and let-7i was increased in AS patients. MEG3 could interact with let-7i in AS fibroblasts, and let-7i overexpression reversed the suppressive effect of MEG3 upregulation on the inflammation and bone formation of AS. Additionally, let-7i could target SOST, and SOST silencing reversed the inhibitory effect of let-7i inhibitor or MEG3 overexpression on the inflammation and bone formation of AS. Furthermore, SOST expression was positively regulated by MEG3, while was negatively regulated by let-7i. Our results revealed that lncRNA MEG3 promoted SOST expression to restrain the progression of AS by sponging let-7i, which provided a treatment target for AS.
摘要：
强直性脊柱炎(AS)是一种以骶髂关节和脊柱附着点炎症为主要特征的疾病。长链非编码RNA（lncRNA）在许多疾病的进展中起着关键作用。然而,关于lncRNA母体表达基因3（MEG3）在AS中的功能的研究很少。定量实时聚合酶链反应（qRT-PCR）用于测量MEG3，microRNAlet-7i，硬化蛋白(SOST),和炎性细胞因子。双荧光素酶报告基因测定,RNA免疫沉淀(RIP)测定和生物素标记的RNA下拉测定用于确认MEG3和let-7i或let-7i和SOST之间的相互作用。此外,进行蛋白质印迹（WB）分析以检测成骨标志物和SOST的蛋白质水平。AS患者MEG3和SOST的表达水平降低，let-7i升高。MEG3可以与AS成纤维细胞中的let-7i相互作用，let-7i过表达逆转了MEG3上调对AS炎症和骨形成的抑制作用。此外,let-7i可以瞄准SOST,SOST沉默逆转了let-7i抑制剂或MEG3过表达对AS炎症和骨形成的抑制作用。此外，SOST表达受MEG3正调控,而let-7i负调控。我们的结果表明，lncRNAMEG3通过海绵let-7i促进SOST表达以抑制AS的进展，这为AS提供了治疗目标。
