Publication Date:
2018-02-01
Description:
Objectives: MicroRNA let -7 family acts as the key regulator of the differentiation of mesenchymal stem cells (MSCs). However, the influence of let -7 b on biological characteristics of stem cells from apical papilla (SCAPs) is still controversial. Materials and methods: In this study, the expression of hsa - let -7 b was obviously downregulated during the osteogenic differentiation of SCAPs. SCAPs were then infected with hsa - let -7 b or hsa - let -7 b inhibitor lentiviruses. The proliferation ability was determined by CCK-8 and flow cytometry. The odonto/osteogenic differentiation capacity was analyzed by alkaline phosphatase (ALP) activity, alizarin red staining, western blot assay, and real-time RT-PCR. Bioinformatics analysis was used to screen out the target of hsa - let -7 b and the target relationship was confirmed by dual luciferase reporter assay. Results: Hsa - let -7 b was of no influence on the proliferation of SCAPs. Interferential expression of hsa - let -7 b increased the ALP activity as well as the formation of calcified nodules of SCAPs. Moreover, the mRNA levels of osteoblastic markers ( ALP , RUNX 2, OSX , OPN , and OCN ) were upregulated while the protein levels of DSPP, ALP, RUNX2, OSX, OPN, and OCN also increased considerably. Conversely, overexpression of hsa - let -7 b inhibited the odonto/osteogenic differentiation capacity of SCAPs. Bioinformatics analysis revealed a putative binding site of hsa - let -7 b in the matrix metalloproteinase 1 ( MMP 1) 3′-untranslated region (3′-UTR). Dual luciferase reporter assay confirmed that hsa - let -7 b targets MMP 1. The odonto/osteogenic differentiation ability of SCAPs ascended after repression of hsa - let -7 b , which was then reversed after co-transfection with si MMP 1. Conclusions: Hsa - let -7 b can suppress the odonto/osteogenic differentiation capacity of SCAPs by targeting MMP 1. This article is protected by copyright. All rights reserved
Electronic ISSN:
0091-7419
Topics:
Biology
,
Chemistry and Pharmacology
,
Medicine
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