Zhu Dongsheng, Zhu Zhitao, Qi Han
Department of Pediatric Surgery, the First People's Hospital of Lianyungang, Affiliated to Xuzhou Medical University, Lianyungang, Jiangsu, 222000, People's Republic of China.
Department of Radiology, the Second People's Hospital of Lianyungang, Lianyungang, Jiangsu, 222000, People's Republic of China.
J Inflamm Res. 2023 Jun 29;16:2675-2687. doi: 10.2147/JIR.S410339. eCollection 2023.
Long non-coding RNA plays an important role in osteogenic differentiation. Nuclear enriched abundant transcript 1 (NEAT1) has been revealed to promote osteogenic differentiation in human bone marrow mesenchymal stem cells (hBMSCs), but the underlying regulatory mechanism remains unknown in acute suppurative osteomyelitis of children.
Osteogenic medium (OM) was used to induce osteogenic differentiation. Quantitative real-time PCR and Western blotting were used to evaluate gene expression. The effects of NEAT1, microRNA 339-5p (miR-339-5p), and salmonella pathogenicity island 1 (SPI1) on osteogenic differentiation were assessed in vitro using alizarin red S staining assays and alkaline phosphatase activity. Interactions between NEAT1, miR-339-5p, and SPI1 were identified using immunoprecipitation, luciferase reporter assays, and chromatin immunoprecipitation.
During osteogenic differentiation, expression of NEAT1 was up-regulated in hBMSCs, and miR-339-5p level was down during osteogenic differentiation. Knockdown of NEAT1 reduced the osteogenic differentiation of hBMSCs, and down-regulation of miR-339-5p may counteract the effect of NEAT1 silencing. SPI1 was a target of miR-339-5p by luciferase reporter assay and was also a transcription factor of NEAT1 by chromatin immunoprecipitation. A positive NEAT1-miR-339-5p-SPI1 feedback loop was found to be present during osteogenic differentiation in hBMSCs.
It was the first study to reveal that the NEAT1-miR-339-5p-SPI1 feedback loop can promote osteogenic differentiation in hBMSCs and shed a new light on the role of NEAT1 during osteogenic differentiation.
长链非编码RNA在成骨分化中起重要作用。核富集丰富转录本1（NEAT1）已被揭示可促进人骨髓间充质干细胞（hBMSCs）的成骨分化，但在儿童急性化脓性骨髓炎中其潜在调控机制仍不清楚。
使用成骨培养基（OM）诱导成骨分化。采用定量实时聚合酶链反应和蛋白质免疫印迹法评估基因表达。使用茜素红S染色试验和碱性磷酸酶活性在体外评估NEAT1、微小RNA 339-5p（miR-339-5p）和沙门氏菌致病岛1（SPI1）对成骨分化的影响。使用免疫沉淀、荧光素酶报告基因检测和染色质免疫沉淀确定NEAT1、miR-339-5p和SPI1之间的相互作用。
在成骨分化过程中，hBMSCs中NEAT1的表达上调，而成骨分化过程中miR-339-5p水平下降。敲低NEAT1可降低hBMSCs的成骨分化，miR-339-5p的下调可能抵消NEAT1沉默的作用。荧光素酶报告基因检测显示SPI1是miR-339-5p的靶标，染色质免疫沉淀显示SPI1也是NEAT1的转录因子。发现hBMSCs成骨分化过程中存在正向的NEAT1-miR-339-5p-SPI1反馈环。
该研究首次揭示NEAT1-miR-339-5p-SPI1反馈环可促进hBMSCs的成骨分化，并为NEAT1在成骨分化中的作用提供了新的线索。
