Department of Outpatient Injection Room Emergency, Weifang People's Hospital, Weifang, Shandong, China.
Eur Rev Med Pharmacol Sci. 2017 Nov;21(21):4777-4783.
Wnt/β-catenin signaling pathway promotes osteoblasts (OBs) differentiation through up-regulating osteoblast-specific gene runt-related transcription factor 2 (RUNX2) expression. It was showed that microRNA-214 (miR-214) was abnormally increased in bone tissue from osteoporosis patients, suggesting its role in osteogenesis. Bioinformatics analysis revealed the complementary binding site between miR-214 and 3'-UTR of β-catenin. This study investigated the effects of miR-214 in regulating β-catenin expression and bone marrow mesenchymal stem cells (BMSCs) differentiating into OB.
BMSCs were induced to differentiate to OB in a specific medium. MiR-214, β-catenin, and RUNX2 expressions were detected. The regulatory relationship between miR-214 and β-catenin was confirmed by dual luciferase reporter gene assay. BMSCs were divided into five groups, including agomir-control, miR-214 agomir, pGPU6-normal control group (pGPU6-NC), pGPU6-β-catenin, and miR-214 agomir + pGPU6-β-catenin groups. β-catenin and RUNX2 levels were tested after 21 days' induction. OB differentiation degree was evaluated by alizarin red staining.
MiR-214 was down-regulated, while β-catenin and RUNX2 were enhanced in the process of BMSCs differentiating into OBs. MiR-214 agomir and/or β-catenin shRNA pGPU6-β-catenin transfection significantly reduced β-catenin expression, declined RUNX2 level, and attenuated OB differentiation in BMSCs.
Wnt/β-catenin signaling pathway was enhanced, while the miR-214 level was decreased in the process of BMSCs differentiating into OBs. Up-regulation of miR-214 inhibited the OB differentiation of BMSCs through targeted suppressing β-catenin expression and attenuating Wnt/β-catenin signaling pathway activity.
Wnt/β-catenin 信号通路通过上调成骨细胞特异性基因 runt 相关转录因子 2（RUNX2）的表达促进成骨细胞（OBs）分化。研究表明，骨质疏松症患者骨组织中 microRNA-214（miR-214）异常升高，提示其在成骨中发挥作用。生物信息学分析显示 miR-214 与β-catenin 3'-UTR 之间存在互补结合位点。本研究探讨了 miR-214 调节β-catenin 表达及骨髓间充质干细胞（BMSCs）向 OB 分化中的作用。
在特定培养基中诱导 BMSCs 向 OB 分化，检测 miR-214、β-catenin 和 RUNX2 的表达。通过双荧光素酶报告基因检测证实 miR-214 与β-catenin 的调控关系。将 BMSCs 分为 agomir-control、miR-214 agomir、pGPU6-正常对照组（pGPU6-NC）、pGPU6-β-catenin 和 miR-214 agomir+pGPU6-β-catenin 组。诱导 21 天后检测β-catenin 和 RUNX2 水平。通过茜素红染色评估 OB 分化程度。
在 BMSCs 向 OB 分化过程中，miR-214 下调，β-catenin 和 RUNX2 上调。miR-214 agomir 和/或β-catenin shRNA pGPU6-β-catenin 转染后，β-catenin 表达显著降低，RUNX2 水平下降，BMSCs 中 OB 分化减弱。
在 BMSCs 向 OB 分化过程中，Wnt/β-catenin 信号通路增强，miR-214 水平降低。上调 miR-214 通过靶向抑制β-catenin 表达和减弱 Wnt/β-catenin 信号通路活性抑制 BMSCs 的 OB 分化。
