Department of Orthopedics and Traumatology, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, P.R. China.
Department of Orthopedics and Traumatology, The First Affiliated Hospital of University of South China, Hengyang, Hunan 421001, P.R. China.
Mol Med Rep. 2020 Nov;22(5):3715-3722. doi: 10.3892/mmr.2020.11489. Epub 2020 Sep 2.
The imbalance induced by inhibition of bone mesenchymal stem cell (BMSC) osteogenic differentiation results in osteoporosis (OP); however, the underlying regulatory mechanism is not completely understood. Long non‑coding RNAs (lncRNAs) serve crucial roles in osteogenic differentiation; therefore, investigating their regulatory role in the process of osteogenic differentiation may identify a promising therapeutic target for OP. The expression of small nucleolar RNA host gene 1 (SNHG1), Dickkopf 1 (DKK1), microRNA (miR)‑101, RUNX family transcription factor 2 (RUNX2), osteopontin (OPN) and osteocalin (OCN) were detected via reverse transcription‑quantitative PCR. The protein expression levels of DKK1, β‑catenin, RUNX2, OPN, OCN, osterix and collagen type I α1 chain were analyzed by performing western blotting. The osteoblastic phenotype was assessed by conducting alkaline phosphatase activity detection and Alizarin Red staining. The interaction between SNHG1 and miR‑101 was validated by bioinformatics and luciferase assays. The regulatory role of SNHG1 in BMSC osteogenic differentiation was assessed. SNHG1 expression was downregulated in a time‑dependent manner during the process of osteogenic differentiation. SNHG1 overexpression inhibited osteogenic differentiation compared with the pcDNA group. The results indicated that SNHG1 and DKK1 directly interacted with miR‑101. Moreover, SNHG1 regulated the Wnt/β‑catenin signaling pathway to inhibit osteogenic differentiation via the miR‑101/DKK1 axis. The present study indicated that lncRNA SNHG1 could attenuate BMSC osteogenic differentiation via the miR‑101/DKK1 axis as a competitive endogenous RNA. Therefore, the present study furthered the current understanding of the potential mechanism underlying lncRNAs in in osteogenic differentiation.
骨间充质干细胞（BMSC）成骨分化抑制导致的失衡会导致骨质疏松症（OP）；然而，其潜在的调控机制尚不完全清楚。长链非编码 RNA（lncRNA）在成骨分化中发挥着至关重要的作用；因此，研究其在成骨分化过程中的调控作用可能为 OP 确定一个有前途的治疗靶点。通过逆转录定量 PCR 检测小核仁 RNA 宿主基因 1（SNHG1）、Dickkopf 1（DKK1）、微小 RNA（miR）-101、RUNX 家族转录因子 2（RUNX2）、骨桥蛋白（OPN）和骨钙素（OCN）的表达。通过蛋白质印迹法分析 DKK1、β-连环蛋白、RUNX2、OPN、OCN、osterix 和胶原 I α1 链的蛋白表达水平。通过碱性磷酸酶活性检测和茜素红染色评估成骨表型。通过生物信息学和荧光素酶测定验证 SNHG1 和 miR-101 之间的相互作用。并评估 SNHG1 在 BMSC 成骨分化中的调节作用。在成骨分化过程中，SNHG1 的表达呈时间依赖性下调。与 pcDNA 组相比，SNHG1 过表达抑制成骨分化。结果表明，SNHG1 和 DKK1 与 miR-101 直接相互作用。此外，SNHG1 通过 miR-101/DKK1 轴调节 Wnt/β-连环蛋白信号通路抑制成骨分化。本研究表明，lncRNA SNHG1 可通过 miR-101/DKK1 轴作为竞争性内源性 RNA 来减弱 BMSC 的成骨分化。因此，本研究进一步加深了对 lncRNA 在成骨分化中潜在机制的理解。
