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| MiR-3609通过靶向调控CCND1抑制成骨细胞分化介导骨质疏松症发生 |
| MiR-3609 mediates osteoporosis by inhibiting differentiation of osteoblast through targeting CCND1 |
| 投稿时间：2023-11-14 修订日期：2024-02-25 |
| DOI： |
| 中文关键词: 骨质疏松症 miR-3609 CCND1 成骨分化 治疗靶点 |
| 英文关键词:osteoporosis miR-3609 CCND1 osteogenesis therapeutic target |
| 基金项目:MiR-3609通过靶向调控CCND1在参与骨质疏松发病的分子机制研究(基金编号:2220004000345) |
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| 中文摘要: |
| 目的 通过生物信息学分析和体外实验结合的方式验证miR-3609在骨质疏松症发病中的作用机制,为骨质疏松症的治疗提供新靶点。方法 通过miRDB、miRWalk、TargetScan三大miRNA靶点分析数据库进行miR-3609下游靶点分析。双荧光素酶实验验miR-3609和下游靶基因之间的靶向关系,RT-qPCR实验和WB实验验证miR-3609对下游靶基因表达的影响及WB实验检测miR-3609对抗凋亡基因Bcl2和成骨相关蛋白Runx2、OPG表达的影响。碱性磷酸酶实验及茜素红实验检测分析miR-3609对成骨细胞成骨分化、矿化的影响。结果 我们通过在线数据库对其相关靶点进行预测,发现CCND1可能是其潜在靶点。双荧光素酶实验验证了二者之间的靶向关系:miR3609的激活会下调成骨细胞中的CCND1表达。碱性磷酸酶和茜素红实验表明,miR3609的激活抑制成骨细胞向成骨分化及矿化。同时,我们发现miR3609的激活下调了成骨细胞成骨相关蛋白Runx2、OPG的表达,同时下调了抗凋亡蛋白Bcl2的表达。结论:通过我们的研究,证实了miR-3609通过靶向抑制CCND1表达抑制成骨细胞成骨分化、矿化,而导致骨质疏松症的发生,靶向抑制miR-3609有望成为骨质疏松症新方向。 |
| 英文摘要: |
| Objective To verify the mechanism of miR-3609 in the pathogenesis of osteoporosis through the combination of bioinformatics analysis and in vitro experiments, providing a new target for the treatment of osteoporosis. Methods The downstream target analysis of miR-3609 was performed using three miRNA target analysis databases, miRDB, miRWalk, and TargetScan. Dual luciferase assay was used to examine the targeting relationship between miR-3609 and downstream target genes, RT-qPCR assay and WB assay to verify the influence of miR-3609 on downstream target gene expression, and WB assay to detect the influence of miR-3609 on anti-apoptosis gene Bcl2 and osteoblast-related proteins Runx2 and OPG expression. The effects of miR-3609 on osteogenic differentiation and mineralization of osteoblasts were analyzed by alkaline phosphatase assay and alizarin red assay. Results We predicted its related targets through online database and find that CCND1 may be a potential target. Dual luciferase assay confirmed the targeting relationship between the two: activation of miR-3609 down-regulates CCND1 expression in osteoblasts. Alkaline phosphatase and alizarin red assay showed that miR-3609 inhibited osteogenic differentiation and mineralization of osteoblasts. At the same time, we found that the activation of miR-3609 down-regulated the expression of osteoblast-related proteins Runx2 and OPG, and down-regulated the expression of anti-apoptotic protein Bcl2. Conclusions Our study confirmed that miR-3609 inhibits osteogenic differentiation and mineralization of osteoblasts through inhibiting the expression of CCND1, leading to the occurrence of osteoporosis. Targeted inhibition of miR-3609 is expected to become a new direction of osteoporosis. |
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