Abstract
Ossification of the posterior longitudinal ligament (OPLL) is a disorder with multiple pathogenic mechanisms and leads to different degrees of neurological symptoms. Recent studies have revealed that non-coding RNA (ncRNA), including long non-coding RNAs (lncRNAs) and microRNAs (miRNAs), could influence the development of OPLL. Nevertheless, the molecular mechanisms linking circular RNAs (circRNAs) and the progression of OPLL is still unknown. The current research explored the expression profiles of OPLL-related circRNAs by microarray analysis, and applied qRT-PCR to validate the results. Subsequently, we confirmed the upregulation of hsa_circ_0007292 in OPLL cells by qRT-PCR and validated the circular characteristic of hsa_circ_0007292 by Sanger sequencing. Fluorescence in situ hybridization (FISH) unveiled that hsa_circ_0007292 was predominantly located in the cytoplasm. Functionally, gain-of-function and loss-of-function experiments showed that hsa_circ_0007292 promoted the osteogenic differentiation of OPLL cells. Mechanistically, the interaction of hsa_circ_0007292 and miR-508-3p was predicted and validated by bioinformatics analysis, dual-luciferase reporter assays, and Ago2 RNA immunoprecipitation (RIP). Similarly, we validated the correlation between miR-508-3p and SATB2. Furthermore, rescue experiments were performed to prove that hsa_circ_0007292 acted as a sponge for miR-508-3p, and SATB2 was revealed to be the target gene of miR-508-3p. In conclusion, our research shows that hsa_circ_0007292 regulates OPLL progression by the miR-508-3p/SATB2 pathway. Our results indicate that hsa_circ_0007292 can be used as a promising therapeutic target for patients with OPLL.
Highlights
Ossification of the posterior longitudinal ligament (OPLL) is a common orthopedic disease, which is characterized by the progressive heterotopic ossification of the posterior longitudinal ligament (PLL) and frequently involves the cervical spinal cord [1]
The function and mechanism of circRNA involved in the development and progression of OPLL are still unknown
We explored the expression profiles of OPLL-related circRNAs by microarray analysis and mainly paid attention to the function and regulatory patterns of hsa_circ_0007292, which exhibited a higher expression in OPLL tissues and cells
Summary
Ossification of the posterior longitudinal ligament (OPLL) is a common orthopedic disease, which is characterized by the progressive heterotopic ossification of the posterior longitudinal ligament (PLL) and frequently involves the cervical spinal cord [1]. OPLL can cause neurological pain and dysfunction by narrowing the vertebral canal and compressing the nerve roots or spinal cord [2]. Once the onset of myelopathic symptoms has started, neurologic function gradually deteriorates, at which point patients require decompressive surgery [3]. The symptoms may reoccur postoperatively because of the gradual progression of ossification over time. It is important to find a way to inhibit the progression of ossified ligaments, www.aging-us.com which could be a better way to treat patients with OPLL
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