Department of Orthopedics, Xuanwu Hospital of Capital Medical University, Beijing 100053, P.R. China.
Department of Orthopedics, Peking University Third Hospital, Beijing 100191, P.R. China.
Int J Mol Med. 2019 May;43(5):2005-2014. doi: 10.3892/ijmm.2019.4130. Epub 2019 Mar 12.
The thoracic ossification of the posterior longitudinal ligament (T‑OPLL) can cause thoracic spinal stenosis, which results in intractable myelopathy and radiculopathy. Our previous whole‑genome sequencing study first reported rs199772854 in the interleukin 17 receptor C (IL17RC) gene as a potentially pathogenic loci for T‑OPLL. The aim of the present study was to examine the effects of the IL17RC gene rs199772854A site mutation on osteogenesis by establishing a model of osteogenic differentiation. IL17RC gene mutation site and wild‑type site mouse embryonic osteoblast (3T3‑E1) models were constructed in order to induce the differentiation of the cells into osteoblasts. Whether the mutation site causes the abnormal expression of the IL17RC gene and osteogenic markers was analyzed by reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) and western blot analysis. The IL17RC gene rs199772854A site mutation was demonstrated to play a biological role through the overexpression of its own gene, and also to significantly increase the expression levels of osteogenic markers. Furthermore, the mutation upregulated the expression of the key proteins, tumor necrosis factor receptor (TNFR)‑associated factor 6 (TRAF6) and nuclear factor (NF)‑κB, in the interleukin (IL)‑17 signaling axis. On the whole, the findings of this study suggest that the IL17RC gene rs199772854A loci mutation propels mouse embryonic osteoblasts towards osteogenic differentiation and may play an important role in the pathogenesis of T‑OPLL. The IL17RC gene may promote osteogenesis through the IL‑17 signaling pathway and may thus be involved in the process of ectopic osteogenesis in T‑OPLL.
胸段后纵韧带骨化(T-OPLL)可导致胸段脊柱狭窄,进而引起难治性脊髓病和神经根病。我们之前的全基因组测序研究首次报道白细胞介素 17 受体 C(IL17RC)基因中的 rs199772854 位点突变为 T-OPLL 的潜在致病基因位点。本研究旨在通过建立成骨分化模型来检测 IL17RC 基因 rs199772854A 位点突变对成骨的影响。构建了 IL17RC 基因突变和野生型位点的小鼠胚胎成骨细胞(3T3-E1)模型,以诱导细胞向成骨细胞分化。通过逆转录-定量聚合酶链反应(RT-qPCR)和 Western blot 分析,分析突变位点是否导致 IL17RC 基因和成骨标志物的异常表达。结果表明,IL17RC 基因 rs199772854A 位点突变通过自身基因的过表达发挥生物学作用,并且显著增加成骨标志物的表达水平。此外,该突变上调了白细胞介素(IL)-17 信号通路中关键蛋白肿瘤坏死因子受体(TNFR)相关因子 6(TRAF6)和核因子(NF)-κB 的表达。总之,本研究结果表明,IL17RC 基因 rs199772854A 位点突变促使小鼠胚胎成骨细胞向成骨分化,可能在 T-OPLL 的发病机制中起重要作用。IL17RC 基因可能通过 IL-17 信号通路促进成骨作用,从而参与 T-OPLL 中的异位成骨过程。