Department of Oral and Maxillofacial Surgery, The Affiliated Hospital of Qingdao University, Qingdao, China.
School of Stomatology of Qingdao University, Qingdao, China.
Oral Dis. 2022 Sep;28(6):1591-1602. doi: 10.1111/odi.13871. Epub 2021 May 25.
Medication-related osteonecrosis of the jaw (MRONJ) is the main adverse side effect of bisphosphonates (BPs), mainly owing to the inhibitory effect of BPs on osteoclastogenesis. CircRNAs were identified to be an important factor in regulating cellular processes. The aim of this study was to explore the effect of mmu_circ_0001066 on BP-inhibited osteoclastogenesis.
The expression of MRONJ-related miRNA in RANKL-induced RAW264.7 cells treated with BP was analyzed using qRT-PCR analysis. Bioinformatics techniques were applied to screen potential circRNAs. Tartrate-resistant acid phosphatase (TRAP) staining and bone resorption assays were used to examine the effect of mmu_circ_0001066 on osteoclastogenesis. Bioinformatics analysis, luciferase reporter assays, and Western blotting assays were performed to investigate the underlying mechanism.
Four MRONJ-related miRNAs were upregulated in BP-treated RAW264.7 cells, and the expression of mmu_circ_0001066 was negatively correlated with those of MRONJ-related miRNAs. Furthermore, the upregulation of mmu_circ_0001066 partially attenuated the inhibitory effect of BP on osteoclastogenesis in RAW264.7 cells. Mechanistically, upregulated miR-16 suppressed osteoclastogenesis and miR-16 inhibitor increased osteoclastogenesis. Furthermore, we have identified that miR-16 is a downstream effector of mmu_circ_0001066.
Our results suggest that mmu_circ_0001066 played an important role in the BP-mediated suppression of osteoclastogenesis, which lays a foundation for identifying mmu_circ_0001066 as a potential biomarker for MRONJ.
药物相关性颌骨坏死（MRONJ）是双膦酸盐（BPs）的主要不良反应，主要归因于 BPs 对破骨细胞生成的抑制作用。CircRNAs 被鉴定为调节细胞过程的重要因素。本研究旨在探讨 mmu_circ_0001066 对 BP 抑制破骨细胞生成的影响。
采用 qRT-PCR 分析分析 RANKL 诱导的 RAW264.7 细胞中 BP 处理后与 MRONJ 相关的 miRNA 的表达。应用生物信息学技术筛选潜在的 circRNAs。采用抗酒石酸酸性磷酸酶（TRAP）染色和骨吸收试验检测 mmu_circ_0001066 对破骨细胞生成的影响。通过生物信息学分析、荧光素酶报告基因检测和 Western blot 检测研究其潜在机制。
BP 处理的 RAW264.7 细胞中 4 种 MRONJ 相关 miRNA 上调，mmu_circ_0001066 的表达与 MRONJ 相关 miRNA 的表达呈负相关。此外，mmu_circ_0001066 的上调部分减弱了 BP 对 RAW264.7 细胞破骨细胞生成的抑制作用。机制上，上调的 miR-16 抑制破骨细胞生成，而 miR-16 抑制剂则增加破骨细胞生成。此外，我们已经确定 miR-16 是 mmu_circ_0001066 的下游效应物。
我们的研究结果表明，mmu_circ_0001066 在 BP 介导的破骨细胞生成抑制中发挥重要作用，为将 mmu_circ_0001066 鉴定为 MRONJ 的潜在生物标志物奠定了基础。
