Zhang Ying, Jia Sansan, Wei Qiushi, Zhuang Zhikun, Li Jitian, Fan Yanan, Zhang Leilei, Hong Zhinan, Ma Xianghao, Sun Ruibo, He Wei, Wang Haibin, Liu Youwen, Li Wuyin
Medical Center of Hip, Luoyang Orthopedic-Traumatological Hospital (Orthopedics Hospital of Henan Province), Luoyang, 471002, Henan, China.
Guangzhou University of Traditional Chinese Medicine, Guangzhou, 510405, Guangdong, China.
Regen Ther. 2020 Dec 28;16:23-31. doi: 10.1016/j.reth.2020.12.003. eCollection 2021 Mar.
We aimed to identify specific circular RNAs (circRNAs) involved in bone repair of trauma-induced osteonecrosis of femoral head (TIONFH) and to explore the potential mechanism. CircRNA sequencing on the blood sample collected from patients with and without TIONFH was performed to select cirRNAs that were significantly differentially expressed, followed by qRT-PCR confirmation. Furthermore, the functions of one selected circRNA and the potential mechanisms in bone repair of TIONFH were validated based on the bone marrow mesenchymal stem cells (BMSCs) and osteoclast-like cells (OLCs) through CCK-8, flow cytometry, transwell assay, luciferase reporter assay, and western blot. A total of 234 upregulated and 148 downregulated differentially expressed circRNAs were identified, and qRT-PCR showed that circRNA_25487 was significantly upregulated in the peripheral blood of TIONFH patients. Luciferase reporter assay confirmed the binding effect between miR-134-3p and circRNA_25487. CircRNA_25487 suppression and miR-134-3p overexpression could promote cell proliferation and invasion while inhibited apoptosis of BMSCs and OLCs. miR-134-3p could target p21. CircRNA_25487 inhibited bone repair in TIONFH by sponging miR-134-3p to upregulate the expression of p21.
我们旨在鉴定参与创伤性股骨头坏死(TIONFH)骨修复的特定环状RNA(circRNA),并探索其潜在机制。对患有和未患有TIONFH的患者采集的血液样本进行circRNA测序,以筛选出差异表达显著的circRNA,随后进行qRT-PCR验证。此外,基于骨髓间充质干细胞(BMSC)和破骨细胞样细胞(OLC),通过CCK-8、流式细胞术、transwell实验、荧光素酶报告基因实验和蛋白质免疫印迹法,验证了一种选定的circRNA的功能及其在TIONFH骨修复中的潜在机制。共鉴定出234个上调和148个下调的差异表达circRNA,qRT-PCR显示circRNA_25487在TIONFH患者外周血中显著上调。荧光素酶报告基因实验证实了miR-134-3p与circRNA_25487之间的结合作用。抑制circRNA_25487和过表达miR-134-3p可促进BMSC和OLC的细胞增殖和侵袭,同时抑制其凋亡。miR-134-3p可靶向p21。circRNA_25487通过海绵吸附miR-134-3p上调p21的表达,从而抑制TIONFH的骨修复。