敲低环状RNA mmu_circ_0001775可提高小鼠骨髓间充质干细胞的成骨能力

Circular RNA mmu_circ_0001775 knockdown improves the osteogenic ability of mouse bone marrow mesenchymal stem cells

背景:近年来利用环状RNA(circRNA)调控干细胞分化成为新的研究热点,将circRNA负载于骨髓间充质干细胞研究其成骨性能,为研发骨缺损修复新型材料具有重要意义。目的:探讨敲低circRNA mmu_circ_0001775对骨髓间充质干细胞成骨能力的影响。方法:生物信息学分析工具推测miR-335-5p关系最为密切的上游circRNA为mmu_circ_0001775,将敲低mmu_circ_0001775的慢病毒、阴性对照病毒转染至骨髓间充质干细胞。qRT-PCR检测各组细胞成骨诱导7,14 d时Runt相关转录因子2、骨钙素、miR-335-5p及mmu_circ_0001775的表达水平,验证其成骨作用,Western blot检测骨形态发生蛋白2、骨桥蛋白的表达。结果与结论:LV-mmu_circ_0001775-BMMSCs组各时间点Runt相关转录因子2、骨钙素表达高于LV-BMMSCs组(P<0.01),骨形态发生蛋白2、骨桥蛋白相对表达量均明显高于LV-BMMSCs组(P<0.05)。结果表明,敲低环状RNA mmu_circ_0001775有利于提高骨髓间充质干细胞的成骨能力。

BACKGROUND:In recent years,the use of circular RNA(circRNA)to regulate stem cell differentiation has become a new research focus.Therefore,loading circRNA on bone marrow mesenchymal stem cells to study their osteogenic properties is of great significance for the development of new materials to repair bone defect OBJECTIVE:To investigate the osteogenic effect of circRNA mmu_circ_0001775 knockdown on bone marrow mesenchymal stem cells.METHODS:Bioinformatics analysis tools speculated that the upstream circRNA most closely related to miR-335-5p was mmu_circ_0001775.Lentivirus and negative control virus knockdown mmu_circ_0001775 were transfected into bone marrow mesenchymal stem cells.qRT-PCR was used to detect the expression levels of Runt-related transcription factor 2,osteocalcin,miR-335-5p,and mmu_circ_0001775 genes at 7 and 14 days of osteogenic induction,respectively,to verify their osteogenic effects.Western blot assay was used to detect the expression levels of bone morphogenetic protein 2 and osteopontin.RESULTS AND CONCLUSION:The expression levels of Runt-related transcription factor 2 and osteocalcin at each time point in LV-mmu_circ_0001775-BMMSCs group were higher than those in LV-BMMSCs group(P<0.01).The relative expression levels of bone morphogenetic protein 2 and osteopontin proteins were significantly higher in the LV-mmu_circ_0001775-BMMSCs group than those of LV-BMMSCs group(P<0.05).The results showed that cirCRNA mmu_circ_0001775 knockdown could enhance the osteogenic ability of bone marrow mesenchymal stem cells.