MicroRNA hsa‐let‐7b suppresses the odonto/osteogenic differentiation capacity of stem cells from apical papilla by targeting MMP1

• Published in: Journal of cellular biochemistry Vol. 119; no. 8; pp. 6545 - 6554
• Main Authors: Wang, Yanqiu, Pang, Xiyao, Wu, Jintao, Jin, Lin, Yu, Yan, Gobin, Romila, Yu, Jinhua
• Format: Journal Article
• Language: English
• Published: United States Wiley Subscription Services, Inc 01.08.2018
• Subjects: 3' Untranslated regions; Adolescent; Adult; Alizarin; Alkaline phosphatase; Antigens, Differentiation - biosynthesis; Antigens, Differentiation - genetics; Assaying; Binding sites; Biocompatibility; Bioinformatics; Cbfa-1 protein; Cell Differentiation; Cholecystokinin; Dental Papilla - cytology; Dental Papilla - metabolism; differentiation; Differentiation (biology); Female; Flow cytometry; hsa‐let‐7b; Humans; Male; Matrix metalloproteinase; matrix metalloproteinase 1; Matrix Metalloproteinase 1 - biosynthesis; Matrix Metalloproteinase 1 - genetics; Mesenchyme; Metalloproteinase; MicroRNAs; MicroRNAs - biosynthesis; MicroRNAs - genetics; miRNA; mRNA; Nodules; Odontogenesis; Osteoblasts; Osteogenesis; Proteins; Ribonucleic acid; RNA; Stem cells; Stem Cells - cytology; Stem Cells - metabolism; stem cells from apical papilla; Transfection; hsa-let-7b; differentiation; stem cells from apical papilla; matrix metalloproteinase 1
• ISSN: 0730-2312; 1097-4644
• DOI: 10.1002/jcb.26737

MicroRNA let‐7 family acts as the key regulator of the differentiation of mesenchymal stem cells (MSCs). However, the influence of let‐7b on biological characteristics of stem cells from apical papilla (SCAPs) is still controversial. In this study, the expression of hsa‐let‐7b was obviously downregulated during the osteogenic differentiation of SCAPs. SCAPs were then infected with hsa‐let‐7b or hsa‐let‐7b 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‐7b and the target relationship was confirmed by dual luciferase reporter assay. Hsa‐let‐7b was of no influence on the proliferation of SCAPs. Interferential expression of hsa‐let‐7b increased the ALP activity as well as the formation of calcified nodules of SCAPs. Moreover, the mRNA levels of osteoblastic markers (ALP, RUNX2, 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‐7b inhibited the odonto/osteogenic differentiation capacity of SCAPs. Bioinformatics analysis revealed a putative binding site of hsa‐let‐7b in the matrix metalloproteinase 1 (MMP1) 3′‐untranslated region (3′‐UTR). Dual luciferase reporter assay confirmed that hsa‐let‐7b targets MMP1. The odonto/osteogenic differentiation ability of SCAPs ascended after repression of hsa‐let‐7b, which was then reversed after co‐transfection with siMMP1. Together, hsa‐let‐7b can suppress the odonto/osteogenic differentiation capacity of SCAPs by targeting MMP1. In this study, alkaline phosphatase activity, alizarin red staining, western blot assay, and real‐time RT‐PCR were used to investigate effects of hsa‐let‐7b/MMP1 axis on the committed differentiation of SCAPs. We found that has‐let‐7b can suppress the odonto/osteogenic differentiation capacity of SCAPs by targeting MMP1.