hsa_circ_0001485 通过靶向 BMPR2 激活 TGFβ-BMP 通路促进成骨分化。
Hsa_circ_0001485 promoted osteogenic differentiation by targeting BMPR2 to activate the TGFβ-BMP pathway.
机构信息
Department of Orthopaedics, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, No. 55 Inner Ring West Road, Guangzhou Higher Education Mega Center, Guangzhou, 510006, Guangdong, China.
Department of Rehabilitation and Recovery, Albury Wodonga Health, Albury, NSW, 2640, Australia.
出版信息
Stem Cell Res Ther. 2022 Sep 5;13(1):453. doi: 10.1186/s13287-022-03150-1.
BACKGROUND
Circular RNAs (circRNAs) are a new type of stable noncoding RNA and have been proven to play a crucial role in osteoporosis. This study explored the role and mechanism of hsa_circ_0001485 in osteogenic differentiation.
METHODS
Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis and Gene Ontology (GO) enrichment analysis were performed according to the previous sequencing data in human bone marrow mesenchymal stem cells (BMSC) before and after the induction of osteogenic differentiation on the differentially expressed circRNAs, to screen out signaling pathways associated with osteogenic differentiation. The hFOB 1.19 cells were used to verify the function and mechanism of specific circRNAs in osteogenic differentiation. Additionally, small interfering fragments and overexpression plasmids were used to determine the role of specific circRNAs during osteogenic differentiation. Furthermore, pull-down experiments and mass spectrometry were performed to determine the proteins that bind to specific circRNAs.
RESULTS
The KEGG and GO enrichment analyses showed that the TGFβ-BMP signaling pathway was related to the osteogenic differentiation process, and four circRNAs were associated with the pathway. The quantitative polymerase chain reaction analysis revealed that hsa_circ_0001485 expression was increased during the osteogenic differentiation process of BMSCs. Knockdown of hsa_circ_0001485 suppressed the activity of the alkaline phosphatase enzyme and the expression of RUNX2, osteopontin, and osteocalcin in the osteogenic hFOB 1.19 cells, whereas overexpression of hsa_circ_0001485 promoted their expression. Additionally, we found that hsa_circ_0001485 and BMPR2 targeted binding to activate the TGFβ-BMP signaling pathway and promoted osteogenic differentiation through mass spectrometry analysis.
CONCLUSION
This study demonstrates that hsa_circ_0001485 is highly expressed in the osteogenic hFOB 1.19 cells, which activate the TGFβ-BMP pathway through targeted binding of BMPR2, and plays a positive role in regulating osteogenic differentiation.
背景
环状 RNA（circRNA）是一种新型稳定的非编码 RNA，已被证明在骨质疏松症中发挥关键作用。本研究探讨了 hsa_circ_0001485 在成骨分化中的作用和机制。
方法
根据人骨髓间充质干细胞（BMSC）在成骨诱导前后差异表达 circRNA 的测序数据，进行京都基因与基因组百科全书（KEGG）分析和基因本体论（GO）富集分析，筛选与成骨分化相关的信号通路。使用 hFOB 1.19 细胞验证特定 circRNA 在成骨分化中的功能和机制。此外，使用小干扰片段和过表达质粒确定特定 circRNA 在成骨分化过程中的作用。进一步通过下拉实验和质谱分析确定与特定 circRNA 结合的蛋白质。
结果
KEGG 和 GO 富集分析表明，TGFβ-BMP 信号通路与成骨分化过程有关，其中 4 个 circRNA 与该通路有关。定量聚合酶链反应分析显示，hsa_circ_0001485 在 BMSC 成骨分化过程中表达增加。hsa_circ_0001485 敲低抑制成骨 hFOB 1.19 细胞碱性磷酸酶活性和 RUNX2、骨桥蛋白和骨钙素的表达，而过表达 hsa_circ_0001485 则促进其表达。此外，我们通过质谱分析发现 hsa_circ_0001485 和 BMPR2 靶向结合激活 TGFβ-BMP 信号通路，通过促进成骨分化。
结论
本研究表明，hsa_circ_0001485 在成骨 hFOB 1.19 细胞中高表达，通过靶向结合 BMPR2 激活 TGFβ-BMP 通路，在调节成骨分化中发挥正向作用。
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