CAJ | 학술논문

目的：探究miR-25在成骨细胞分化过程中的作用及其作用机制。方法β-甘油磷酸（β-glycerophos-phate，β-GP）和抗坏血酸（ ascorbic acid）诱导成骨细胞矿化，茜素红（ Alizarin Red S）染色实验观察成骨细胞的矿化情况。生物信息学运算预测miR-25的靶基因。实时定量PCR法检测miR-25以及其他相关基因的表达。过表达实验和抑制实验用于研究miR-25在成骨细胞分化中的作用及其与靶基因之间的相互关系。荧光素酶报告基因实验验证miR-25与其靶基因之间的结合。结果 miR-25的表达随成骨分化过程而逐渐升高。过表达miR-25通过转录后调控其靶基因TWIST1的表达来促进成骨分化。作为调控成骨细胞分化的的转录因子，扭曲基础螺旋—环—螺旋蛋白1（ TWIST1）是miR-25的靶基因。结论 miR-125通过作用于靶基因 TWIST1在成骨细胞的分化中发挥了重要的调控作用，表明调控miR-25的表达，可能成为骨代谢疾病新的治疗靶点。
목적：탐구miR-25재성골세포분화과정중적작용급기작용궤제。방법β-감유린산（β-glycerophos-phate，β-GP）화항배혈산（ ascorbic acid）유도성골세포광화，천소홍（ Alizarin Red S）염색실험관찰성골세포적광화정황。생물신식학운산예측miR-25적파기인。실시정량PCR법검측miR-25이급기타상관기인적표체。과표체실험화억제실험용우연구miR-25재성골세포분화중적작용급기여파기인지간적상호관계。형광소매보고기인실험험증miR-25여기파기인지간적결합。결과 miR-25적표체수성골분화과정이축점승고。과표체miR-25통과전록후조공기파기인TWIST1적표체래촉진성골분화。작위조공성골세포분화적적전록인자，뉴곡기출라선—배—라선단백1（ TWIST1）시miR-25적파기인。결론 miR-125통과작용우파기인 TWIST1재성골세포적분화중발휘료중요적조공작용，표명조공miR-25적표체，가능성위골대사질병신적치료파점。
[Abstrast] Objective To explore the role of miR-25 in the process of osteoblast differentiation and its mechanism. Me thods β-glycerophosphate (βglycerophosphate,β-GP) and ascorbic acid ( ascorbic acid) induced osteoblast mineraliza-tion, Alizarin Red ( Alizarin Red S) experiment were performed to observe stained cells into bone mineralization.Bioinformat-ics computing predicted target genes of miR-25.Real-time quantitative PCR was used to detect the expression of miR-25 and other related genes.Overexpression experiments and inhibitions experiments were used to investigate the relationship between of miR-25 in osteoblast differentiation and its relationship with target gene.Luciferase gene reporter experiments were used to demonstrate miR-25 binding to its target genes.Results The expression of miR-25 increased gradually in the process of osteo-genic differentiation. Overexpression of miR-25 promotes osteogenic differentiation through post transcriptional regulate TWIST1 expression of its target genes .As a transcription factor that regulates the differentiation of bone cells ,TWIST1 is a target gene of miR-25.Conclusion miR-25 plays an important role in the differentiation of osteoblasts by acting on target gene TWIST1, which indicates that the expression of miR-25 may be a new therapeutic target for bone metabolism.