Osteogenic Differentiation

SIPA1L1[Circular RNA]

miR-204-5p[miRNA]

ALPL[mRNA]

stem cells from apical papilla (SCAPs)

Homo sapiens (human)

Stem Cell Res Ther. 2020 Nov 2;11(1):461. doi: 10.1186/s13287-020-01970-7.

Circular RNA SIPA1L1 regulates osteoblastic differentiation of stem cells from apical papilla via miR-204-5p/ALPL pathway.

CCK-8 assay;Dual-luciferase reporter assay;Flow cytometry assay;Western blot;Flow Cytometry assay;Luciferase reporter assay;

BACKGROUND: Osteogenesis is a complex biological process which requires the coordination of multiple molecular mechanisms. This research aimed to explore the biological role and underlying regulatory mechanism of circSIPA1L1 during the osteogenic differentiation of stem cells from apical papilla (SCAPs). METHODS: EdU retention assay, flow cytometry assay, and CCK-8 assay were used to evaluate the proliferation capacity of SCAPs. Western blot assay, alkaline phosphatase (ALP), and alizarin red staining (ARS) were conducted to investigate the biological roles of circSIPA1L1 and miR-204-5p. Fluorescence in situ hybridization was applied for circSIPA1L1 localization. Dual-luciferase reporter assay was performed to prove the interaction of circSIPA1L1 and miR-204-5p. RESULTS: CircSIPA1L1 had no significant effect on the proliferative capacity of SCAPs. CircSIPA1L1 promotes osteogenic differentiation of SCAPs by serving as a miRNA sponge for miR-204-5p. Either knockdown of circSIPA1L1 or overexpression of miR-204-5p significantly suppresses osteogenic differentiation of SCAPs. CONCLUSIONS: CircSIPA1L1 upregulates ALPL through targeting miR-204-5p and promotes the osteogenic differentiation of SCAPs.