Down‐regulation of hsa‐circ‐0107593 promotes osteogenic differentiation of hADSCs via miR‐20a‐5p/SMAD6 signaling

Abstract

ObjectivesIncreasing evidence indicated circRNAs were involved in stem cells osteogenesis differentiation. Herein, we aimed to clarify the role of hsa‐circ‐0107593 during the osteogenesis process of human adipose‐derived stem cells (hADSCs) and the underlying mechanisms.MethodsThe ring structure of hsa‐circ‐0107593 was confirmed using RNase R treatment and Sanger sequencing. Nucleoplasmic separation and fluorescence in situ hybridization detected hsa‐circ‐0107593 distribution. Lentivirus and siRNA were used to modulate the expression of hsa‐circ‐0107593, and the binding relationship between hsa‐circ‐0107593 and miR‐20a‐5p was verified by luciferase assay and RNA immunoprecipitation. We detected the osteogenic activity of hADSCs through alkaline phosphatase staining, alizarin red S staining, real‐time polymerase chain reaction (RT‐PCR), western blot, and cellular immunofluorescence experiment. In vivo, micro‐computed tomography was performed to analyze bone formation around skull defect.ResultsRT‐PCR results exhibited that hsa‐circ‐0107593 was downregulated while miR‐20a‐5p was upregulated during hADSCs osteogenesis. In vivo and in vitro experiments results indicated that knocking down hsa‐circ‐0107593 promoted the osteogenic differentiation of hADSCs, while overexpression of hsa‐circ‐0107593 showed an inhibitory effect on hADSCs osteogenic differentiation. In vitro experiment results showed hsa‐circ‐0107593 acted as a hADSCs osteogenic differentiation negative factor for it inhibited the suppressing effect of miR‐20a‐5p on SMAD6.ConclusionKnocking down hsa‐circ‐0107593 acts as a positive factor of the osteogenic differentiation of hADSCs via miR‐20a‐5p/SMAD6 signaling.

Comprehensive Analysis of Long Non-coding RNA,miRNA，and mRNA-associated Competitive Endogenous RNA Networks in the Peripheral Blood of Preterm Infants with Bronchopulmonary Dysplasia

“…Studies have shown that downregulation of hsa-circ-0107593 promotes the osteogenic differentiation of hADSCs through miR-20a-5p/SMAD6 signaling (51). Atorvastatin may be involved in lipid-lowering therapy by downregulating microRNA-20a-5p (52).…”

Exploring the Role of Circular RNA in Bone Biology: A Comprehensive Review

“…Regulates the switch between osteogenesis and adipogenesis by sponging miR-338-3p [57] circ-3626 Promotes osteogenesis by via miR-338-3p/Runx2 axis [7] Circ-FK501 Promotes osteogenic differentiation via microRNA-205-5p/RUNX2 axis [58] hsa-circ-0107593 Inhibits osteogenic differentiation via miR-20a-5p/SMAD6 signaling [59] Circ_0006873 Inhibits osteogenic differentiation via miR-20a-/SMURF2 signaling [60] Circ_0114581 Promotes osteogenic differentiation via the MiR-155-5p/HNRNPA3 axis [61] Circ_0001825 Promotes osteogenesis via miR-1270/SMAD5 axis [62] Circ_0036872 Promotes osteogenesis via miR-143-3p/IGF2 axis [63] CircZNF367 Inhibits osteogenic differentiation via reducing HuR-mediated mRNA stability of LRP5 [64] Circ_C4orf36 Promotes osteogenesis by regulating VEGFA [65] Circ-Sirt1 Promotes osteogenesis by activating Sirt1 and Wnt/β-catenin pathway [66] CircRNAs also play roles in maintaining bone homeostasis by regulating bone metabolism and the remodeling processes. For example, circRNA_ circ_0006859 was found to inhibit osteogenesis and promote adipogenesis in human in human bone marrow mesenchymal stem cells by targeting the signaling miR-431-5p/Rho-associated protein kinase 1 (ROCK1) [67] and circRNA-CDR1 promotes adipogenesis and inhibits osteogenic differentiation in osteonecrosis of the femoral head [68].…”

Post-Transcriptional Regulatory Crosstalk between MicroRNAs and Canonical TGF-β/BMP Signalling Cascades on Osteoblast Lineage: A Comprehensive Review

“…MiR-20a, a mechanosensitive miRNA was found to enhance fluid shear stress-mediated osteogenic differentiation of rat BMSCs via the activation of BMP-2 signalling pathway by targeting Bambi and Smad6 [ 270 ]. In addition, silencing of hsa-circ-0107593 was found to upregulate miR-20a-5p expression, in which stimulated osteogenic differentiation of human ADSCs through targeting of Smad6 [ 271 ].…”