Department of Orthopedics, The Second Hospital of Shanxi Medical University, Taiyuan, 030001 Shanxi, China.
Biomed Res Int. 2022 Aug 26;2022:1624446. doi: 10.1155/2022/1624446. eCollection 2022.
To probe into the effect of LncRNA TUG1 on the healing of closed tibial fracture in mice.
The closed tibial fracture model of mice was established, selecting the mouse osteoblast line MC3T3-E1, with the cells separated into four groups. The expression levels of TUG1 and miR-221-3p were determined by RT-qPCR analysis, with the targeting relationship between TUG1 and miR-221-3p authenticated by dual luciferase reporter (DLR) assay, detection of cell migration (CM) ability based on Transwell cell migration (TCM) assay, and cell proliferation (CP) acquired by cell counting kit-8 (CCK-8).
Prediction results of the target gene by bioinformatics software showed that miR-221-3p had binding sites with the 3'-UTR of TUG1, and DLR assay authenticated the targeting relationship between LncRNA TUG1 and miR-221-3p. Downregulation of TUG1 inhibited osteoblast CP and CM and promoted osteoblast cell apoptosis (CA). Cell cycle analysis indicated that miR-221-3p provoked cell cycle arrest in G1 stage of MC3T3-E1 cells. The siLncRNA-NC group had higher anticyclin D1 and D3 levels than the siLncRNA TUG1 group, with a lower CA rate in the former, implying that miR-221-3p overexpression inhibited osteoblast CP and CM and LncRNA TUG1 inhibited CA. Downregulation of miR-221-3p partly reversed the retardation out of downregulating TUG1 on osteoblast CP and CM. Bcl-2 level was higher in the LncRNA TUG1 group compared to the siLncRNA TUG1 and miR-221-3p overexpression groups, with remarkably lower SDF-1 level in the miR-221-3p overexpression group than those in the control, miRNA-NC, and LncRNA TUG1 groups.
The downregulation of miR-221-3p by LncRNA TUG1 can promote the healing of closed tibial fractures in mice.
探讨 LncRNA TUG1 对小鼠闭合性胫骨骨折愈合的影响。
构建小鼠闭合性胫骨骨折模型，选择小鼠成骨细胞系 MC3T3-E1，将细胞分为四组。通过 RT-qPCR 分析测定 TUG1 和 miR-221-3p 的表达水平，通过双荧光素酶报告（DLR）测定验证 TUG1 与 miR-221-3p 的靶向关系，通过 Transwell 细胞迁移（TCM）测定检测细胞迁移（CM）能力，通过细胞计数试剂盒-8（CCK-8）检测细胞增殖（CP）。
生物信息学软件预测靶基因结果表明，miR-221-3p 与 TUG1 的 3'-UTR 具有结合位点，DLR 测定验证了 LncRNA TUG1 与 miR-221-3p 的靶向关系。下调 TUG1 抑制成骨细胞 CP 和 CM，并促进成骨细胞细胞凋亡（CA）。细胞周期分析表明，miR-221-3p 可引起 MC3T3-E1 细胞周期停滞在 G1 期。siLncRNA-NC 组的 cyclin D1 和 D3 水平高于 siLncRNA TUG1 组，前者的 CA 率较低，表明 miR-221-3p 过表达抑制成骨细胞 CP 和 CM，LncRNA TUG1 抑制 CA。下调 miR-221-3p 部分逆转了下调 TUG1 对成骨细胞 CP 和 CM 的抑制作用。LncRNA TUG1 组的 Bcl-2 水平高于 siLncRNA TUG1 组和 miR-221-3p 过表达组，miR-221-3p 过表达组的 SDF-1 水平明显低于对照组、miRNA-NC 组和 LncRNA TUG1 组。
LncRNA TUG1 通过下调 miR-221-3p 可促进小鼠闭合性胫骨骨折的愈合。
