Down-regulation of Noggin and miR-138 coordinately promote osteogenesis of mesenchymal stem cells

• Published in: Journal of molecular histology 2017-12, Vol.48 (5-6), p.427-436
• Main Authors: Sun, Xing-Kun, Zhou, Jin, Zhang, Lei, Ma, Tian, Wang, Yu-Han, Yang, Yan-Mei, Tang, Yan-Ting, Li, Hong, Wang, Li-Jun
• Format: Article
• Language: English
• Subjects: Alkaline phosphatase; Alkaline Phosphatase - metabolism; Animals; Biomedical and Life Sciences; Biomedicine; Carrier Proteins - metabolism; Cbfa-1 protein; Cell Biology; Cell Differentiation - genetics; Cell Proliferation; Developmental Biology; Down-Regulation - genetics; Extracellular Signal-Regulated MAP Kinases - metabolism; Flow cytometry; Gene expression; Immunophenotyping; Life Sciences; Mesenchymal stem cells; Mesenchymal Stromal Cells - cytology; Mesenchymal Stromal Cells - enzymology; Mesenchymal Stromal Cells - metabolism; Mesenchyme; Mice; MicroRNAs - genetics; MicroRNAs - metabolism; Noggin protein; Nucleotides; Oligonucleotides; Oligonucleotides - metabolism; Original Paper; Osteocytes; Osteogenesis; Osteogenesis - genetics; Polymerase chain reaction; Rehabilitation; RNA, Small Interfering - metabolism; Signal Transduction; siRNA; Smad protein; Smad Proteins - metabolism; Stem cell transplantation; Stem cells
• ISSN: 1567-2379; 1567-2387
• DOI: 10.1007/s10735-017-9740-5

Mesenchymal stem cells (MSCs) can differentiate to osteocytes under suitable conditions. In recent years, micro-nucleotides have been progressively used to modulate gene expression in cells due to the consideration of safety. Our present study aimed to investigate whether co-delivery of Noggin-siRNA and antimiR-138 enhances the osteogenic effect of MSCs. Using a murine MSC line, C3H/10T1/2 cells, the delivery efficiency of Noggin-siRNA and antimiR-138 into MSCs was evaluated by quantitative real-time polymerase chain reaction (qRT-PCR). Cell phenotype and proliferation capacity was assessed by flow cytometry and MTT assay respectively. The osteogenesis of MSCs was tested by Alkaline Phosphatase (ALP) staining, qRT-PCR, and western blot analyses. Our results demonstrated that the expression of Noggin and miR-138 were significantly silenced in MSCs by Noggin-siRNA and/or antimiR-138 delivery, while the phenotype and proliferation capacity of MSCs were not affected. Down-regulation of Noggin and miR-138 cooperatively promoted osteogenic differentiation of MSCs. The ALP positive cells reached about 83.57 ± 10.18%. Compared with single delivery, the expression of osteogenic related genes, such as Alp, Col-1, Bmp2, Ocn and Runx2, were the highest in cells with co-delivery of the two oligonucleotides. Moreover, the protein level of RUNX2, and the ratios of pSMAD1/5/SMAD1/5 and pERK1/2/ERK1/2 were significantly increased. The activation of Smad, Erk signaling may constitute the underlying mechanism of the enhanced osteogenesis process. Taken together, our study provides a safe strategy for the clinical rehabilitation application of MSCs in skeletal deficiency.