The osteogenic differentiation of mesenchymal stem cells (MSCs) has potential clinical values in the treatment of bone-related diseases. Long non-coding RNA H19 and microRNA-140-5p (miR-140-5p) have attracted much attention of researchers by virtue of their biological importance in cell differentiation and bone formation. Moreover, bioinformatics analyses suggest that miR-140-5p have the potential to bind with H19 and SATB homeobox 2 (SATB2). In this study, we further explored whether H19 could regulate osteogenic differentiation of human bone marrow-derived MSCs (BM-MSCs) by miR-140-5p/SATB2 axis. RT-qPCR assay was conducted to examine the expression of H19, miR-140-5p and SATB2. The osteogenic differentiation capacity of BM-MSCs was assessed through alkaline phosphatase (ALP) activity and osteogenic marker expression. The relationships among H19, miR-140-5p and SATB2 were examined through bioinformatics analyses, luciferase reporter assay, RIP assay and RNA pull-down assay. H19 expression was remarkably increased and miR-140-5p expression was dramatically reduced during osteogenic differentiation of BM-MSCs. Functional analyses revealed that H19 overexpression or miR-140-5p depletion accelerated osteogenic differentiation of BM-MSCs. Conversely, H19 loss or miR-140-5p increase suppressed osteogenic differentiation of BM-MSCs. MiR-140-5p was confirmed as a target of H19, and miR-140-5p could bind to SATB2 as well. Moreover, H19 knockdown reduced SATB2 expression by upregulating miR-140-5p. Additionally, miR-140-5p depletion antagonized the inhibitory effect of H19 knockdown on osteogenic differentiation of BM-MSCs. And, miR-140-5p inhibited osteogenic differentiation of BM-MSCs by targeting SATB2. In conclusion, H19 promoted osteogenic differentiation of BM-MSCs through regulating miR-140-5p/SATB2 axis, deepening our understanding on the molecular mechanisms of H19 in coordinating osteogenesis.

中文翻译：

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长链非编码RNA H19通过调节microRNA-140-5p/SATB2轴促进人骨髓间充质干细胞成骨分化

间充质干细胞 (MSCs) 的成骨分化在治疗骨相关疾病方面具有潜在的临床价值。长链非编码 RNA H19 和 microRNA-140-5p (miR-140-5p) 因其在细胞分化和骨形成中的生物学重要性而备受研究人员的关注。此外，生物信息学分析表明 miR-140-5p 有可能与 H19 和 SATB 同源框 2 (SATB2) 结合。在本研究中，我们进一步探讨了 H19 是否可以通过 miR-140-5p/SATB2 轴调节人骨髓源性 MSCs (BM-MSCs) 的成骨分化。进行RT-qPCR测定以检查H19、miR-140-5p和SATB2的表达。BM-MSCs 的成骨分化能力通过碱性磷酸酶 (ALP) 活性和成骨标志物表达进行评估。H19之间的关系，通过生物信息学分析、荧光素酶报告基因分析、RIP 分析和 RNA 下拉分析检测 miR-140-5p 和 SATB2。在BM-MSCs的成骨分化过程中，H19表达显着增加，miR-140-5p表达显着降低。功能分析表明，H19 过表达或 miR-140-5p 耗竭加速了 BM-MSCs 的成骨分化。相反，H19 缺失或 miR-140-5p 增加抑制了 BM-MSC 的成骨分化。MiR-140-5p 被确认为 H19 的靶点，并且 miR-140-5p 也可以与 SATB2 结合。此外，H19 敲低通过上调 miR-140-5p 降低了SATB2 表达。此外，miR-140-5p 耗竭拮抗了 H19 敲低对 BM-MSCs 成骨分化的抑制作用。和，miR-140-5p 通过靶向 SATB2 抑制 BM-MSCs 的成骨分化。总之，H19通过调节miR-140-5p/SATB2轴促进BM-MSCs的成骨分化，加深了我们对H19协调成骨分子机制的理解。