环状 RNA SIPA1L1 通过 miR-204-5p/ALPL 通路调控根尖乳头干细胞成骨分化。
Circular RNA SIPA1L1 regulates osteoblastic differentiation of stem cells from apical papilla via miR-204-5p/ALPL pathway.
机构信息
Key Laboratory of Oral Diseases of Jiangsu Province and Stomatological Institute of Nanjing Medical University, 136 Hanzhong Road, Nanjing, 210029, Jiangsu, China.
Endodontic Department, School of Stomatology, Nanjing Medical University, Nanjing, Jiangsu, China.
出版信息
Stem Cell Res Ther. 2020 Nov 2;11(1):461. doi: 10.1186/s13287-020-01970-7.
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.
背景
成骨作用是一个复杂的生物学过程,需要多个分子机制的协调。本研究旨在探讨 circSIPA1L1 在根尖乳头干细胞(SCAPs)成骨分化过程中的生物学作用及其潜在的调控机制。
方法
采用 EdU 保持试验、流式细胞术分析和 CCK-8 检测评估 SCAPs 的增殖能力。通过 Western blot 分析、碱性磷酸酶(ALP)和茜素红染色(ARS)实验研究 circSIPA1L1 和 miR-204-5p 的生物学作用。应用荧光原位杂交检测 circSIPA1L1 的定位。双荧光素酶报告基因实验验证 circSIPA1L1 和 miR-204-5p 的相互作用。
结果
circSIPA1L1 对 SCAPs 的增殖能力没有显著影响。circSIPA1L1 通过作为 miR-204-5p 的 miRNA 海绵促进 SCAPs 的成骨分化。circSIPA1L1 的敲低或 miR-204-5p 的过表达均显著抑制 SCAPs 的成骨分化。
结论
circSIPA1L1 通过靶向 miR-204-5p 上调 ALPL,促进 SCAPs 的成骨分化。
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