Zhang Lei, Xu Yidan, Liu Ningning, Xu Mengjie, Tuerxun Refeina, Shahatibieke Mireyeti, Yao Yuan
Department of Prosthodontics, Stomatology Research Institute of Xinjiang Uygur Autonomous Region, The First Affiliated Hospital of Xinjiang Medical University, Affiliated Stomatological Hospital of Xinjiang Medical University, Urumqi, Xinjiang, People's Republic of China.
Xinjiang West Cell Biotechnology Co. LTD, Urumqi, Xinjiang, People's Republic of China.
Sci Rep. 2025 Jun 6;15(1):19950. doi: 10.1038/s41598-025-03711-3.
Circular RNA (circRNA) is a unique closed ring structure of noncoding RNA. Although many human diseases have been confirmed to be inextricably linked with circRNAs, whether circRNAs have a potential regulatory function in the osteogenic differentiation of bone marrow mesenchymal stem cells (BMMSCs) remains to be further elucidated. In a previous study by our team, all the differentially expressed circRNAs and messenger RNAs (mRNAs) involved in the osteogenic differentiation of BMMSCs were identified via high-throughput sequencing, and a competing endogenous RNA (ceRNA) regulatory network was constructed via bioinformatics analysis. The circRNA_1809/miR-370-3p/Kitlg axis may be involved in regulating the osteogenic differentiation of BMMSCs. In this study, gene knockdown/overexpression, small interfering RNA (siRNA) transfection and mimic/inhibitor treatment were used to evaluate the regulatory effects of circRNA_1809 on the miR-370-3p/Kitlg pathway and the osteogenic differentiation of rat bone marrow mesenchymal stem cells (rBMMSCs) in vitro. The results revealed that circRNA_1809 was upregulated and that miR-370-3p was downregulated during the osteogenic differentiation of rBMMSCs. The low expression of circRNA_1809 significantly downregulated the expression of Kitlg and decreased the protein expression of ALP and RUNX2. The expression of miR-370-3p was negatively correlated with the expression of Kitlg in rBMMSCs and the ability to undergo osteogenic differentiation. In addition, a dual-luciferase assay confirmed the binding of miR-370-3p and circRNA_1809, and si-circRNA_1809 + miR-370-3p inhibitor cotransfection reversed some of the downregulation of Kitlg induced by si-circRNA_1809, whereas si-circRNA_1809 + miR-370-3p mimic increased the downregulation of Kitlg. Therefore, circRNA_1809 may promote the expression of Kitlg by regulating miR-370-3p and subsequently promote the osteogenic differentiation of rBMMSCs.
环状RNA（circRNA）是一种独特的非编码RNA闭环结构。尽管许多人类疾病已被证实与circRNA有着千丝万缕的联系，但circRNA在骨髓间充质干细胞（BMMSC）成骨分化中是否具有潜在调节功能仍有待进一步阐明。在我们团队之前的一项研究中，通过高通量测序鉴定了参与BMMSC成骨分化的所有差异表达的circRNA和信使RNA（mRNA），并通过生物信息学分析构建了竞争性内源RNA（ceRNA）调控网络。circRNA_1809/miR-370-3p/Kitlg轴可能参与调节BMMSC的成骨分化。在本研究中，采用基因敲低/过表达、小干扰RNA（siRNA）转染和模拟物/抑制剂处理，在体外评估circRNA_1809对miR-370-3p/Kitlg通路及大鼠骨髓间充质干细胞（rBMMSC）成骨分化的调节作用。结果显示，在rBMMSC成骨分化过程中circRNA_1809表达上调，而miR-370-3p表达下调。circRNA_1809低表达显著下调Kitlg表达，并降低碱性磷酸酶（ALP）和 runt相关转录因子2（RUNX2）的蛋白表达。在rBMMSC中，miR-370-3p的表达与Kitlg的表达以及成骨分化能力呈负相关。此外，双荧光素酶报告基因检测证实了miR-370-3p与circRNA_1809的结合，并且si-circRNA_1809与miR-370-3p抑制剂共转染逆转了si-circRNA_1809诱导的Kitlg部分下调，而si-circRNA_1809与miR-370-3p模拟物共转染则增加了Kitlg的下调。因此，circRNA_1809可能通过调节miR-370-3p促进Kitlg的表达，进而促进rBMMSC的成骨分化。
