Department of Orthodontics, Guangzhou Women and Children's Medical Center, Guangzhou, Guangdong, 510000, China.
Department of Stomatology, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong, 510080, China; Guangdong Key Laboratory of Stomatology, Sun Yat-sen University, Guangzhou, Guangdong, 510080, China.
Differentiation. 2020 Nov-Dec;116:1-8. doi: 10.1016/j.diff.2020.09.005. Epub 2020 Oct 9.
Osteogenic differentiation of dental pulp stem cells (DPSCs) is considered as a promising strategy in posterior maxilla tooth implantation. Information on the function and mechanisms of long non-coding RNAs (lncRNAs) in osteogenic differentiation of DPSCs is growing, however, the mechanism of LINC00968 and miR-3658 in regulating osteogenic differentiation of DPSCs still needs to be explored. In this study, the LINC00968 and miR-3658 expression level was upregulated and downregulated in DPSCs and peri-implantitis DPSCs (pDPSCs) treated with bone morphogenic protein (BMP)2, respectively. Moreover, the effects of LINC00968 and miR-3658 on BMP2-induced osteogenic differentiation of DPSCs in vitro using Alizarin Red S staining, alkaline phosphatase (ALP) activity, quantitative real time PCR and Western blot assays showed that overexpression of LINC00968 significantly promoted mineralized bone matrix, alkaline phosphatase (ALP), runt-related transcription factor 2 (RUNX2), and osterix (OSX) expression levels for osteogenic differentiation of DPSCs and pDPSCs; and overexpression of miR-3658 showed an opposite result that inhibited osteogenic differentiation of DPSCs and pDPSCs. Luciferase reporter assay showed that luciferase activities of LINC00968-WT reporter and RUNX2-WT reporter were strongly suppressed by miR-3658 overexpression. In addition, the miR-3658 upregulation interfered ectopic bone formation in vivo stimulated by LINC00968. In general, we had identified a novel molecular pathway involving LINC00968/miR-3658/RUNX2 during DPSCs and pDPSCs differentiation into osteoblasts, which might facilitate bone anabolism.
牙髓干细胞(DPSCs)的成骨分化被认为是上颌后牙种植的一种有前途的策略。关于长非编码 RNA(lncRNA)在 DPSCs 成骨分化中的功能和机制的信息正在不断增加,然而,LINC00968 和 miR-3658 在调节 DPSCs 成骨分化中的机制仍有待探索。在这项研究中,分别上调和下调了 DPSCs 和骨形成蛋白 2(BMP2)处理的牙周炎牙髓干细胞(pDPSCs)中的 LINC00968 和 miR-3658 的表达水平。此外,通过茜素红 S 染色、碱性磷酸酶(ALP)活性、实时定量 PCR 和 Western blot 分析,研究了 LINC00968 和 miR-3658 对 DPSCs 体外 BMP2 诱导的成骨分化的影响,结果表明,LINC00968 的过表达显著促进了 DPSCs 和 pDPSCs 的矿化骨基质、碱性磷酸酶(ALP)、成 Runt 相关转录因子 2(RUNX2)和骨形成蛋白 2(OSX)的表达水平,促进了 DPSCs 和 pDPSCs 的成骨分化;而 miR-3658 的过表达则呈现出相反的结果,抑制了 DPSCs 和 pDPSCs 的成骨分化。荧光素酶报告基因实验表明,miR-3658 过表达强烈抑制了 LINC00968-WT 报告基因和 RUNX2-WT 报告基因的荧光素酶活性。此外,miR-3658 的上调干扰了 LINC00968 体内刺激的异位骨形成。总之,我们已经确定了一个新的分子途径,涉及 LINC00968/miR-3658/RUNX2 在 DPSCs 和 pDPSCs 向成骨细胞分化过程中,这可能有助于骨合成代谢。