Department of Orthopedics, Chifeng Municipal Hospital, No.1 Zhaowuda Road, Chifeng, 024000, Inner Mongolia, People's Republic of China.
In Vitro Cell Dev Biol Anim. 2020 Jan;56(1):49-58. doi: 10.1007/s11626-019-00405-9. Epub 2019 Nov 15.
Previous research manifested that miR-140-3p was a latent biomarker for osteoporosis. Nevertheless, the mechanism of miR-140-3p in osteoporosis is still not clear and needs ulteriorly studying. The purpose of our paper was to ulteriorly probe the underlying mechanism of miR-140-3p on osteoporosis. Firstly, based on the data acquired from GEO database, we found that miR-140-3p was highly expressed; meanwhile, MCF2L was lowly expressed in osteoporosis patients. Upregulation/downregulation of miR-140-3p by miR-140-3p mimic/inhibitor restrained/promoted MC3T3-E1 cell viability and differentiation. However, miR-140-3p over-expression/downregulation accelerated/repressed MC3T3-E1 cell apoptosis. MCF2L was forecasted as a target of miR-140-3p by miRanda, miRWalk, and TargetScan miRNA target gene prediction software. Luciferase reporter assay confirmed that MCF2L could be directly targeted by miR-140-3p. Moreover, we identified that the expression of MCF2L was negatively regulated by miR-140-3p. From rescue assays, we discovered that knockdown of MCF2L weakened the promoting influence of miR-140-3p ablation on MC3T3-E1 cell viability and differentiation, and receded the suppressing impact of miR-140-3p reduction on MC3T3-E1 cell apoptosis. Above all, this research disclosed that miR-140-3p repressed preosteoblast viability and differentiation while promoted preosteoblast apoptosis via targeting MCF2L. Our discoveries might afford a theoretical basis of developing a latent novel target for osteoporosis therapy.
先前的研究表明，miR-140-3p 是骨质疏松症的潜在生物标志物。然而，miR-140-3p 在骨质疏松症中的作用机制尚不清楚，需要进一步研究。本文旨在进一步探讨 miR-140-3p 对骨质疏松症的潜在作用机制。首先，基于 GEO 数据库获得的数据，我们发现 miR-140-3p 高表达，同时骨质疏松症患者 MCF2L 低表达。miR-140-3p 模拟物/抑制剂的上调/下调抑制/促进了 MC3T3-E1 细胞的活力和分化。然而，miR-140-3p 的过表达/下调加速/抑制了 MC3T3-E1 细胞的凋亡。miRanda、miRWalk 和 TargetScan miRNA 靶基因预测软件预测 MCF2L 是 miR-140-3p 的靶基因。荧光素酶报告基因实验证实 MCF2L 可被 miR-140-3p 直接靶向。此外，我们发现 MCF2L 的表达受 miR-140-3p 的负调控。通过挽救实验，我们发现 MCF2L 的敲低减弱了 miR-140-3p 缺失对 MC3T3-E1 细胞活力和分化的促进作用，并减弱了 miR-140-3p 减少对 MC3T3-E1 细胞凋亡的抑制作用。综上所述，本研究表明 miR-140-3p 通过靶向 MCF2L 抑制成骨前体细胞活力和分化，促进成骨前体细胞凋亡。我们的发现可能为骨质疏松症治疗提供新的潜在靶点的理论依据。
