miR-3614-5p prevents endothelial differentiation and angiogenesis in human exfoliated deciduous teeth by targeting lysophosphatidic acid receptor 2
摘要
The contribution of microRNAs (miRNAs) to the endothelial differentiation of stem cells from human exfoliated deciduous teeth (SHED) remains largely unclear. This study sought to uncover novel miRNAs implicated in this process. SHED were obtained from deciduous teeth and verified via flow cytometric profiling of CD34, CD90, and CD105. Cells were induced toward endothelial lineage and subjected to miRNA sequencing at 0, 7, and 14 d. Differentially expressed miRNAs were validated by quantitative Polymerase Chain Reaction (qPCR). The influence of miR-3614-5p on endothelial differentiation was examined using qPCR, Western blotting, immunofluorescence, and angiogenesis assays. Direct targeting of lysophosphatidic acid receptor 2 (LPAR2) was tested through luciferase reporter assays, and rescue experiments were performed by LPAR2 overexpression. Results showed that SHED expressed CD34 (19.5%), CD90 (99.1%), and CD105 (92.5%), confirming effective isolation. Sequencing identified 9 consistently downregulated and 11 consistently upregulated miRNAs at both days 7 and 14, with miR-3614-5p denoting stable upregulation and qPCR confirmation. Functionally, miR-3614-5p exerted opposing effects: its overexpression suppressed von Willebrand factor and CD31 expression and impaired tube formation, whereas its inhibition enhanced marker expression and angiogenesis. Mechanistically, LPAR2 was validated as a direct target of miR-3614-5p. Importantly, LPAR2 overexpression reversed the inhibitory effects of miR-3614-5p on endothelial differentiation and angiogenesis. Together, these findings indicated that miR-3614-5p negatively regulates SHED endothelial differentiation and angiogenesis by directly targeting LPAR2, highlighting its potential role as a molecular regulator in this process.