Department of Orthopedics, The Second Affiliated Hospital of Soochow University, 1055 Sanxiang Road, Suzhou, 215004, Jiangsu, China.
Department of Orthopedics, The First People's Hospital of Nantong, Nantong, 226001, Jiangsu Province, China.
J Orthop Surg Res. 2020 Jul 10;15(1):255. doi: 10.1186/s13018-020-01780-w.
The dysregulation of proliferation and apoptosis plays a significant role in the pathogenesis of postmenopausal osteoporosis (PO). MicroRNAs play an important role in regulating apoptosis of MC3T3-E1 cells. However, the role and potential mechanism of miR-708 for regulating HO-induced apoptosis is unknown. This study aimed to investigate the protective function of miR-708 in HO-induced apoptosis of MC3T3-E1 osteoblasts.
MC3T3-E1 was co-cultured with HO for 8 h, then, flow cytometry, malondialdehyde (MDA), and glutathione peroxidase (Gpx) levels were measured to establish the oxidative model. MiRNA microarray was performed to assess differentially expressed miRNAs between control and HO-treated MC3T3-E1 cells. We then performed RT-PCR to identify the relative expression of miR-708 and PTEN. After transfected MC3T3-E1 with miR-708 mimics, flow cytometry, MDA, and Gpx level were performed to identify the apoptosis rate and oxidative stress in these groups. Furthermore, we small interfering RNA of PTEN to identify the role of PTEN in HO-induced apoptosis of MC3T3-E1 cells.
HO (100 nM) could significantly induce the apoptosis of MC3T3-E1 cells. Moreover, HO could significantly increase the MDA level and downregulated Gpx level. RT-PCR found that HO significantly decrease the level of miR-708. Compared with HO group, HO + miR-708 mimic significantly decreased the apoptosis rate.
miR-708 plays a protective role in HO-induced MC3T3-E1 osteoblasts apoptosis and its protective effect is proceeded by regulating ROS level and PTEN expression level.
细胞增殖和凋亡失调在绝经后骨质疏松症（PO）的发病机制中起重要作用。MicroRNAs 在调节 MC3T3-E1 细胞凋亡中起重要作用。然而，miR-708 调节 HO 诱导的细胞凋亡的作用及其潜在机制尚不清楚。本研究旨在探讨 miR-708 在 HO 诱导的 MC3T3-E1 成骨细胞凋亡中的保护作用。
将 MC3T3-E1 与 HO 共培养 8 小时，然后通过流式细胞术、丙二醛（MDA）和谷胱甘肽过氧化物酶（Gpx）水平来建立氧化模型。进行 miRNA 微阵列分析以评估对照和 HO 处理的 MC3T3-E1 细胞之间差异表达的 miRNAs。然后，我们进行 RT-PCR 以鉴定 miR-708 和 PTEN 的相对表达。转染 MC3T3-E1 后，用 miR-708 模拟物进行流式细胞术、MDA 和 Gpx 水平，以鉴定这些组中的细胞凋亡率和氧化应激。此外，我们用 PTEN 的小干扰 RNA 鉴定了 PTEN 在 HO 诱导的 MC3T3-E1 细胞凋亡中的作用。
HO（100 nM）可显著诱导 MC3T3-E1 细胞凋亡。此外，HO 可显著增加 MDA 水平并下调 Gpx 水平。RT-PCR 发现 HO 显著降低了 miR-708 的水平。与 HO 组相比，HO + miR-708 模拟物可显著降低细胞凋亡率。
miR-708 在 HO 诱导的 MC3T3-E1 成骨细胞凋亡中发挥保护作用，其保护作用是通过调节 ROS 水平和 PTEN 表达水平来实现的。
