Circ_0087960 stabilizes KDM5B by reducing SKP2 mediated ubiquitination degradation and promotes osteogenic differentiation in periodontal ligament stem cells

Abstract

Background: Periodontitis is a common chronic oral disease among the world. Periodontal ligament stem cells (PDLSCs) has been proved to be a promising tool for the treatment of periodontitis due to their capability of generating periodontal tissues. Circ_0087960 and KDM5B have been shown to participate in the process of osteogenic differentiation with unclear function and mechanism. Methods: Circ_0087960 and KDM5B expressions were detected during the osteogenic induction of PDLSCs. The functions of circ_0087960 and KDM5B were validated by manipulating their expression with shRNA. ChIP and luciferase reporter assays were used to prove the KDM5B-based osteogenic gene regulation. Co-IP assay was used to determine the interaction between SKP2 and KDM5B. In vivo ubiquitination assay was used to test the modification of KDM5B by SKP2. RNA pull-down was used to demonstrate the interaction between circ_0087960 and KDM5B. Results: Circ_0087960 and KDM5B were found to be upregulated in the osteogenic differentiation of PDLSCs and promote the expression of related genes. KDM5B could directly bind and promote the expression of Runx2, ALP and OCN. KDM5B protein level in PDLSCs was controlled by SKP2-mediated protein ubiquitination and degradation. Circ_0087960 was identified to bind to KDM5B protein and protect it against SKP2-induced protein degradation, leading to the upregulation of osteogenic genes. Conclusion: Circ_0087960 and KDM5B could be applied as promising therapeutic methods to stimulate the osteogenic differentiation of PDLSCs, expanding their capability in the treatment of periodontitis.

Keywords

• Circ_0087960
• KDM5B
• SKP2
• Osteogenic induction
• PDLSCs