Parathyroid hormone (PTH) acts as a regulator of calcium homeostasis and bone remodeling. Runx2, an essential transcription factor in bone, is required for osteoblast differentiation. Noncoding RNAs such as long noncoding RNAs (lncRNAs) and microRNAs (miRNAs) play crucial roles in regulating gene expression in osteoblasts. In this study, we investigated the effects of PTH on osteoblast differentiation via Runx2, lncRNA, and miRNA expression in human bone marrow stromal cells (hBMSCs) and human osteoblastic cells (MG63). PTH-treatment of hBMSCs for 24 h, 7 days, and 14 days stimulated Runx2 mRNA expression. Using bioinformatics tools, we identified 17 lncRNAs originating from human Runx2 gene. Among these, lnc-SUPT3H-1:16 (RUNX2-AS1:32) expression was highly up-regulated by the 7 d PTH-treatment in hBMSCs. We also identified miR-6797-5p as the putative target of lnc-SUPT3H-1:16 and Runx2 using bioinformatics tools. PTH-treatment increased the expression of miR-6797-5p in hBMSCs, and overexpression of miR-6797-5p decreased osteoblast differentiation in MG63 cells, suggesting a role for lnc-SUPT3H-1:16 as sponge molecule. A luciferase gene reporter assay identified direct targeting of miR-6797-5p with lnc-SUPT3H-1:16 and 3′UTR Runx2 in MG63 cells. Thus, PTH stimulated the expression of lnc-SUPT3H-1:16, miR-6797-5p and Runx2, and due to the sponging mechanism of lnc- SUPT3H-1:16 towards miR-6797-5p, Runx2 was protected, resulting in the promotion of osteoblast differentiation.

甲状旁腺激素（PTH）充当钙稳态和骨骼重塑的调节剂。Runx2是骨骼中必不可少的转录因子，是成骨细胞分化所必需的。诸如长非编码RNA（lncRNA）和微RNA（miRNA）之类的非编码RNA在调节成骨细胞中的基因表达中起着至关重要的作用。在这项研究中，我们研究了PTH通过Runx2，lncRNA和miRNA在人骨髓基质细胞（hBMSCs）和人成骨细胞（MG63）中表达对成骨细胞分化的影响。hBMSC的PTH处理24小时，7天和14天可刺激Runx2 mRNA表达。使用生物信息学工具，我们鉴定了来自人类Runx2基因的17个lncRNA。其中，lnc-SUPT3H-1：16（RUNX2-AS1：32）表达在hBMSC中受7 d PTH处理高度上调。我们还使用生物信息学工具将miR-6797-5p确定为lnc-SUPT3H-1：16和Runx2的假定靶标。PTH处理增加了hBMSCs中miR-6797-5p的表达，miR-6797-5p的过表达降低了MG63细胞的成骨细胞分化，提示lnc-SUPT3H-1：16作为海绵分子起作用。萤光素酶基因报告基因分析鉴定了在MG63细胞中，miR-6797-5p被lnc-SUPT3H-1：16和3'UTR Runx2直接靶向。因此，PTH刺激了lnc-SUPT3H-1：16，miR-6797-5p和Runx2的表达，并且由于lnc-SUPT3H-1：16对miR-6797-5p的海绵化作用，Runx2得到了保护，从而导致促进成骨细胞分化。miR-6797-5p的过表达和过表达降低了MG63细胞的成骨细胞分化，表明lnc-SUPT3H-1：16作为海绵分子起作用。萤光素酶基因报告基因分析鉴定了在MG63细胞中，miR-6797-5p被lnc-SUPT3H-1：16和3'UTR Runx2直接靶向。因此，PTH刺激了lnc-SUPT3H-1：16，miR-6797-5p和Runx2的表达，并且由于lnc-SUPT3H-1：16对miR-6797-5p的海绵化作用，Runx2得到了保护，从而导致促进成骨细胞分化。miR-6797-5p的过表达和过表达降低了MG63细胞的成骨细胞分化，表明lnc-SUPT3H-1：16作为海绵分子起作用。萤光素酶基因报告基因分析鉴定了在MG63细胞中，miR-6797-5p被lnc-SUPT3H-1：16和3'UTR Runx2直接靶向。因此，PTH刺激了lnc-SUPT3H-1：16，miR-6797-5p和Runx2的表达，并且由于lnc-SUPT3H-1：16朝向miR-6797-5p的海绵化机制，Runx2得到了保护，从而导致促进成骨细胞分化。