Spry1在miR-21调控人骨髓间充质干细胞成骨分化中的作用

Role of Spry1 in osteogenic differentiation of human bone marrow mesenchymal stem cells under miR-21 regulation

背景:前期研究发现,miR-21在骨髓间充质干细胞成骨分化中表达上升,但是miR-21在骨髓间充质干细胞成骨分化中的作用及分子机制尚不明确。目的:验证miR-21的靶基因Spry1,探明Spry1在人骨髓间充质干细胞成骨分化中的作用。方法:荧光素酶报告验证miR-21靶基因Spry1,Western Blot检测Spry1在人骨髓间充质干细胞成骨过程的表达。构建Spry1慢病毒表达载体,感染人骨髓间充质干细胞。碱性磷酸酶、茜素红染色、RT-PCR和Western Blot分析Spry1高表达后人骨髓间充质干细胞成骨能力的改变。结果与结论:荧光素酶报告提示Spry1为miR-21的靶基因,在人骨髓间充质干细胞成骨过程中表达量下降。上调Spry1能够抑制人骨髓间充质干细胞的成骨分化。结果表明Spry1作为miR-21的靶基因负向调控人骨髓间充质干细胞的成骨分化,在骨形成过程发挥着重要作用。

BACKGROUND：Previous studies have found that miR-21 expression is increased during osteogenic differentiation of bone marrow mesenchymal stem cells, but the action and molecular mechanism of miR-21 are stil unclear. OBJECTIVE：To verify the target gene of miR-21, Spry1, and to explore the role of Spry1 in osteogenic differentiation of human bone marrow mesenchymal stem cells. METHODS：Luciferase report was used to verify Spry1 gene targeted by miR-21, and western blot assay was used to detect the expression of Spry1 in the osteogenesis of human bone marrow mesenchymal stem cells. Spry1 expression vector was established and transfected into human bone marrow mesenchymal stem cells. Osteogenesis ability of human bone marrow mesenchymal stem cells was analyzed after Spry1 high expression by alkaline phosphatase, alizarin red staining, RT-PCR and western blot. RESULTS AND CONCLUSION：Luciferase report suggested that Spry1 was a target gene of miR-21. The expression level of Spry1 was decreased in the osteogenesis of human bone marrow mesenchymal stem cells. Increasing expression of Spry1 could inhibit osteogenic differentiation of human bone marrow mesenchymal stem cells. These results indicate that Spry1 as a target gene of miR-21 negatively regulates osteogenic differentiation of human bone marrow mesenchymal stem cells, and plays an important role in bone formation process.