miR-155-5p调控破骨细胞分化的体外研究

miR-155-5p regulates osteoclast differentiation in chronic apical periodontitis.

目的:探究miR-155-5p在破骨细胞分化中的作用。方法:将巨噬细胞集落刺激因子(macrophage colony-stimulating factor,M-CSF)和核因子κB受体活化素配体(receptor activator of nuclear factor kappa-B ligand,RANKL)加入RAW264.7细胞中诱导其向破骨细胞分化,达指定天数后,TRAP染色观察破骨细胞形成并计数,实时荧光定量PCR(qRT-PCR)检测破骨细胞分化相关基因mRNA表达水平,同时检测miR-155-5p的表达。过表达和抑制miR-155-5p后,诱导RAW264.7细胞分化,TRAP染色观察破骨细胞形成,qRT-PCR检测miR-155-5p和破骨细胞分化相关基因的mRNA表达水平。结果:miR-155-5p在破骨细胞分化过程中逐步下调。过表达miR-155-5p抑制破骨细胞分化,抑制miR-155-5p促进破骨细胞分化。结论:miR-155-5p调控破骨细胞分化。

Objective:To investigate the role of miR-155-5p during osteoclast differentiation.Methods:Murine RAW264.7 cells were conducted by seeding cells on six-well plates in osteoclast differentiation medium which consisted of 10%FBS,supplemented with 50 ng/mL recombinant mouse-receptor activator of nuclear factor kappa-B ligand(RANKL)and 30 ng/mL recombinant mouse-macrophage colony-stimulating factor(M-CSF).Differentiated cells were harvested at designated times for TRAP staining and total RNA extraction.TRAP staining was performed to detect the formation of osteoclasts and count the osteoclast number.Real-time quantitative PCR(qRT-PCR)was used to detect the expression level of miR-155-5p and osteoclast differentiation-related genes.After overexpression and downregulation of miR-155-5p,RAW264.7 cells were induced by M-CSF and RANKL.Then TRAP staining and qRT-PCR were performed to detect the formation of osteoclasts and the expression level of osteoclast differentiation-related genes.Results:MiR-155-5p was gradually down-regulated during osteoclast differentiation.Over-expression of miR-155-5p could inhibit osteoclast differentiation,while suppression of miR-155-5p did the opposite effect.Conclusion:MiR-155-5p regulated the osteoclast differentiation process.