Department of Neurosurgery, Third Affiliated Hospital of Soochow University, Changzhou City, 213003, Jiangsu, China.
Department of Geriatrics, Third Affiliated Hospital of Soochow University, Changzhou City, 213003, Jiangsu, China.
Stem Cell Res Ther. 2021 Mar 1;12(1):157. doi: 10.1186/s13287-021-02214-y.
As one of the most common chronic diseases in the world, osteoporosis occurs especially in postmenopausal women. Circular RNAs (circRNAs) are emerging as major drivers in human disease. The aim of the present study was to analyse circRNA expression profiles in osteoporosis and to explore the clinical significance and the regulatory molecular mechanism of hsa_circ_0006859 during osteoporosis.
Exosomes were isolated from clinically collected serum samples. A circRNA microarray was performed to screen differentially expressed circRNAs. Quantitative real-time PCR (qRT-PCR) and western blot were performed to analyse target gene mRNA expression and protein expression. Alizarin red staining (ARS) was performed to evaluate the mineralization ability of human bone marrow mesenchymal stem cells (hBMSCs). Oil Red O staining was performed to evaluate the lipid droplet formation ability of hBMSCs. Bioinformatics analysis and the luciferase reporter assay were performed to investigate the interaction between two genes.
Hsa_circ_0006859 was identified as one of the most upregulated circRNAs in the microarray analysis. Hsa_circ_0006859 in exosomes was upregulated in osteoporosis patients compared to healthy controls. Hsa_circ_0006859 differentiated osteopenia or osteoporosis patients from healthy controls with high sensitivity and specificity. Hsa_circ_0006859 suppressed osteoblastic differentiation and promoted adipogenic differentiation of hBMSCs. Hsa_circ_0006859 directly bound to miR-431-5p, and ROCK1 was identified as a novel target gene of miR-431-5p. Hsa_circ_0006859 is a competing endogenous RNA (ceRNA) of miR-431-5p that promotes ROCK1 expression. Hsa_circ_0006859 suppressed osteogenesis and promoted adipogenesis by sponging miR-431-5p to upregulate ROCK1.
Exosomal hsa_circ_0006859 is a potential biomarker for postmenopausal osteoporosis and controls the balance between osteogenesis and adipogenesis in hBMSCs by sponging miR-431-5p.
骨质疏松症是世界上最常见的慢性疾病之一，尤其发生在绝经后妇女中。环状 RNA（circRNA）作为人类疾病的主要驱动因素而崭露头角。本研究旨在分析骨质疏松症中的 circRNA 表达谱，并探讨 hsa_circ_0006859 在骨质疏松症中的临床意义和调控分子机制。
从临床收集的血清样本中分离出外泌体。进行 circRNA 微阵列筛选差异表达的 circRNA。通过定量实时 PCR（qRT-PCR）和 Western blot 分析靶基因 mRNA 表达和蛋白表达。进行茜素红染色（ARS）评估人骨髓间充质干细胞（hBMSC）的矿化能力。进行油红 O 染色评估 hBMSC 的脂滴形成能力。通过生物信息学分析和荧光素酶报告基因检测进行两个基因相互作用的研究。
hsa_circ_0006859 在微阵列分析中被鉴定为最上调的 circRNA 之一。与健康对照组相比，骨质疏松症患者的外泌体中 hsa_circ_0006859 上调。hsa_circ_0006859 能够以高灵敏度和特异性区分骨质疏松症患者和健康对照者。hsa_circ_0006859 抑制成骨细胞分化，促进 hBMSC 向脂肪细胞分化。hsa_circ_0006859 直接与 miR-431-5p 结合，ROCK1 被鉴定为 miR-431-5p 的新靶基因。hsa_circ_0006859 是 miR-431-5p 的竞争性内源 RNA（ceRNA），通过海绵吸附 miR-431-5p 促进 ROCK1 表达。hsa_circ_0006859 通过海绵吸附 miR-431-5p 抑制成骨分化，促进脂肪生成，从而调控 hBMSC 中骨形成和脂肪形成的平衡。
外泌体 hsa_circ_0006859 是绝经后骨质疏松症的潜在生物标志物，通过海绵吸附 miR-431-5p 调控 hBMSC 中骨形成和脂肪形成的平衡。
