miR-31促进骨髓间充质干细胞的增殖和迁移

miR-31 promotes the proliferation and migration of bone marrow mesenchymal stem cells

背景:骨髓间充质干细胞已被广泛应用于临床治疗神经退行性相关疾病,但由于细胞在受损伤部位的存活率和迁移率低,从而降低了其疗效。目的:探讨miR-31是否能提高骨髓间充质干细胞的迁移和增殖能力。方法:培养和鉴定C57BL/6小鼠骨髓间充质干细胞,将细胞分为对照组、miR-31 agomir组和miR-31 antagomir组。将第3代骨髓间充质干细胞接种于6孔板(1×105/孔),待细胞达到50%-80%融合度,将miR-31 agomir和miR-31 antagomir用不含血清的DMEM/F12培养基稀释后添加到6孔板,转染24 h后,用CCK-8分析细胞的增殖水平以及Transwell实验分析细胞的迁移能力,Western blot检测基质金属蛋白酶2和CXC趋化因子受体4的蛋白表达。结果与结论:①在不加转染试剂的情况下将miR-31成功转染至骨髓间充质干细胞,并发出红色荧光;②转染后,与对照组相比,miR-31 agomir组细胞的增殖能力增强,与时间呈正比(P<0.05);与对照组相比,miR-31 agomir组细胞的迁移能力增强(P<0.05),基质金属蛋白酶2和CXC趋化因子受体4蛋白表达上调(P<0.05);③结果表明,miR-31可以提高骨髓间充质干细胞的增殖能力,并且促进骨髓间充质干细胞的迁移,这为间充质干细胞高效靶向迁移至损伤部位奠定了基础。

BACKGROUND:Bone marrow mesenchymal stem cells have been widely used in the clinical treatment of neurodegenerative-related diseases,but their efficacy is reduced by low cell survival and migration rates at the site of injury.OBJECTIVE:To investigate whether miR-31 can enhance the migration and proliferation of bone marrow mesenchymal stem cells.METHODS:Bone marrow mesenchymal stem cells from C57BL/6 mice were cultured and identified,and the cells were divided into control,miR-31 agomir,and miR-31 antagomir groups.Bone marrow mesenchymal stem cells at passage 3 were inoculated in six-well plates(1×105/well).When the cells reached 50%-80%fusion,miR-31 agomir and miR-31 antagomir were added to the six-well plates after dilution with serum-free DMEM/F12.After 24 hours of transfection,the proliferation level of cells was analyzed by CCK-8 assay as well as the migration ability of cells was analyzed by Transwell assay.Protein expression of matrix metalloproteinase 2 and CXC chemokine receptor 4 was detected by western blot assay.RESULTS AND CONCLUSION:(1)miR-31 was successfully transfected with bone marrow mesenchymal stem cells without transfection reagents and emitted red fluorescence.(2)After transfection,the proliferation ability of cells in the miR-31 agomir group was enhanced compared with the control group,which increased proportionally with time(P<0.05).Compared with the control group,miR-31 promoted the migratory ability of bone marrow mesenchymal stem cells in the miR-31 agomir group(P<0.05)and also upregulated protein expression of matrix metalloproteinase 2 and CXC chemokine receptor 4(P<0.05).(3)The results indicated that miR-31 could improve the proliferation ability of bone marrow mesenchymal stem cells and promote the migration of bone marrow mesenchymal stem cells,which provides a basic study for efficient targeting of mesenchymal stem cell migration to the site of injury.