Tianjin Medical University, Tianjin, China; Department of Spinal Surgery, Tianjin Union Medical Center, Tianjin, China.
Department of Spinal Surgery, Tianjin Union Medical Center, Tianjin, China.
Biochem Biophys Res Commun. 2020 Apr 30;525(2):498-504. doi: 10.1016/j.bbrc.2020.02.126. Epub 2020 Feb 25.
Many studies had identified that MicroRNAs (miRNAs) could affect bone metabolism by regulating the expression of various proteins. This study explored the effect and mechanism of miR-532-3p on osteogenic differentiation.
We analyzed the content of miR-532-3p in osteoporosis patients, osteoporosis rats, and osteogenic induced MC3T3-E1 cells. MiR-532-3p mimic or inhibitor utilized to alter intracellular miR-532-3p content. MTT method executed to detect the effect of miR-532-3p on osteoblast proliferation. Real-time qPCR, Western blot, alkaline phosphatase staining, and alizarin red staining utilized to ascertain the influence of miR-532-3p on osteogenic differentiation. Then, databases and a dual-luciferase reporter gene assay used to verify the target of miR-532-3p. Furthermore, the lentiviral vector was utilized to overexpress interesting target gene expression and checked whether the target gene was involved in the regulation of osteogenic differentiation by miR-532-3p.
MiR-532-3p expression boosted in low bone mineral density (BMD) patients and rats. In MC3T3-E1 cells, miR-532-3p expression gradually decreased as osteogenic induction matures. MiR-532-3p mimic negatively regulated succinate dehydrogenase (SDH) activity, alkaline phosphatase (ALP) activity, mineralization ability, the osteogenic-associated gene (Col1A1, Runx2, ALP, OPN, and OCN) and E-26 transformation specific-1 (ETS1) expression of MC3T3-E1 cells. Things are the opposite of the miR-532-3p inhibitor. ETS1 identified as the miR-532-3p target gene, and miR-532-3p could inhibit its expression. Besides, improved ETS1 expression could rescue the suppressive effect of miR-532-3p mimic on osteogenic differentiation.
miR-532-3p can suppress osteogenic differentiation by downregulating ETS1 expression.
许多研究已经表明，MicroRNAs（miRNAs）可以通过调节各种蛋白质的表达来影响骨代谢。本研究探讨了 miR-532-3p 对成骨分化的影响及其机制。
我们分析了骨质疏松症患者、骨质疏松症大鼠和成骨诱导的 MC3T3-E1 细胞中 miR-532-3p 的含量。利用 miR-532-3p 模拟物或抑制剂来改变细胞内 miR-532-3p 的含量。通过 MTT 法检测 miR-532-3p 对成骨细胞增殖的影响。利用实时 qPCR、Western blot、碱性磷酸酶染色和茜素红染色来确定 miR-532-3p 对成骨分化的影响。然后，利用数据库和双荧光素酶报告基因实验来验证 miR-532-3p 的靶基因。此外，利用慢病毒载体过表达感兴趣的靶基因，并检测靶基因是否参与 miR-532-3p 对成骨分化的调节。
miR-532-3p 在低骨密度（BMD）患者和大鼠中表达增加。在 MC3T3-E1 细胞中，随着成骨诱导的成熟，miR-532-3p 的表达逐渐降低。miR-532-3p 模拟物负调控琥珀酸脱氢酶（SDH）活性、碱性磷酸酶（ALP）活性、矿化能力、成骨相关基因（Col1A1、Runx2、ALP、OPN 和 OCN）和 E-26 转化特异性-1（ETS1）的表达。miR-532-3p 抑制剂的情况则相反。ETS1 被鉴定为 miR-532-3p 的靶基因，miR-532-3p 可以抑制其表达。此外，提高 ETS1 的表达可以挽救 miR-532-3p 模拟物对成骨分化的抑制作用。
miR-532-3p 可以通过下调 ETS1 的表达来抑制成骨分化。
