Objective: To explore the role of cathepsin K (CTSK) regulated by mir-30a-wnt/β-catenin signaling pathway in cementogenic differentiation of periodontal ligament stem cells (PDLSCs).
Methods: Human PDLSCs isolated by limiting dilution culture were induced by enamel matrix protein derivative (EMD) for differentiation into cementoblast-like cells. MicroRNA chip technique was employed to screen the differentially expressed microRNAs in the cells during induced differentiation. The effect of inhibiting miR-30a on CTSK expression in the induced cells was examined using RT-PCR and Western blotting. Ceramic scaffolds coated with PDLSCs treated with EMD and transfected with the miR-30a inhibitor or a lentiviral vector for CTSK overexpression were prepared and implanted subcutaneously in nude mice, and 8 weeks later the cellular expressions of cementoblast markers CAP and CEMP-1 were detected with immunohistochemistry to verify whether CTSK participate in cementogenic differentiation of PDLSCs. The role of wnt signaling pathway in miR-30a-mediated regulation of CTSK expression was explored by examining CTSK protein expressions after blocking wnt signaling in PDLSCs.
Results: In PDLSCs with EMD-induced differentiation into cementoblast-like cells, multiple microRNAs exhibited differential expressions; and among them, miR-30a was specifically and significantly up-regulated ( < 0.05). Up-regulation of miR-30a obviously increased the expression of CTSK ( < 0.05) and promoted PDLSCs to form cementum-like tissues with high expressions of CAP and CEMP-1. The regulatory effect of miR-30a on CTSK expression was obviously attenuated after inhibiting wnt/β-catenin signaling pathway.
Conclusion: EMD induces cementogenic differentiation of PDLSCs possibly by up-regulating the expression of miR-30a, which further activates the wnt/β-catenin signaling pathway to enhance the expression of CTSK.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8586862 | PMC |
| http://dx.doi.org/10.12122/j.issn.1673-4254.2021.10.01 | DOI Listing |
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