Abstract

 
 
 
 Purpose: To determine whether miR-452 regulates osteoblast differentiation (OD) in human periodontal ligament stem cells (hPDLSCs) by targeting polycomb-group protein BMI1.
 Methods: hPDLSCs were stimulated to differentiate upon treatment with mineralization liquid. Quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting were used to measure mRNA and protein expressions, respectively. Alkaline phosphatase (ALP) activity and Alizarin red staining were used to determine the osteogenic differentiation (OD) of hPDLSCs. The bioinformatics software, Targetscan, was used to predict the potential target of miR-452, while luciferase assay, qRT- PCR, and western blot were employed to verify the target gene of miR-452, BMI1.
 Results: MiR-452 was downregulated during the OD of hPDLSCs, but miR-452 overexpression inhibited the OD of hPDLSCs. BMI1 was identified as a direct target gene of miR-452 during the OD of hPDLSCs, while miR-452 overexpression correlated inversely with BMI1 expression during OD of hPDLSCs.
 Conclusion: Overexpression of miR-452 suppresses the OD of hPDLSCs by targeting BMI1.This study may provide potential diagnostic and therapeutic basis for OD in hPDLSCs.
 
 
 
Highlights
Periodontal tissue damage causes oral organ defects and has adverse effects on health for many people [1]
MiR-452 expression decreased in Human periodontal ligament stem cells (hPDLSCs) during osteogenic differentiation (OD)
To investigate the effects of miR-452 on OD of hPDLSCs, hPDLSCs were stably transfected with LV-miR-452. Quantitative real-time polymerase chain reaction (qRT-PCR) showed that hPDLSCs transfected with LV-miR-452 expressed higher levels of miR-452 than that transfected with LV-negative controls (NC) (p < 0.01; Figure 2 A), indicating the successful transfection
Summary
Periodontal tissue damage causes oral organ defects and has adverse effects on health for many people [1]. The miR-452 mimic (miR-452), negative control (NC) of miR-452 mimic (miR-NC), miR-452 inhibitor (miR-452 inh), NC of inhibitor (miR-NC inh), and the short hairpin (shRNA) sequences targeting the BMI1 gene (5'-AAGGAGGAGGT GAATGATAAA-3'), LNA-miR-452 (50 nM) and LNA-miR-NC (50 nM) were transfected into hPDLSCs using Lipofectamine 2000. This assay was used to detect mineralization nodules in hPDLSCs. Briefly, hPDLSCs were washed (PBS), fixed with paraformaldehyde (4 % (w/v), 30 min, at room temperature (RT)), stained with Alizarin red (30 min, at RT), and washed (PBS) for three times. The Dunnett multiple comparison test was applied to identify differences between groups
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
