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Differentiation of a Human Neural Stem Cell Line on Three Dimensional Cultures, Analysis of MicroRNA and Putative Target Genes
Published on: April 12, 2015
LncRNA MEG3 promotes osteogenesis of hBMSCs by regulating miR-21-5p / SOD3 axis
Shuangli Wang1, Gaoxin Xiong2, Rende Ning2
11Department of Bone and Joint Surgery, Institute of Orthopedic Diseases, the First Affiliated Hospital, Jinan University, Guangzhou, China; 2Department of Joint Surgery, the Third Affiliated Hospital, Anhui Medical University, Hefei, China.
Background:
This study aimed to investigate the role of long non-coding (Lnc) RNA MEG3 on the osteogenesis of human bone marrow mesenchymal stem cells (hBMSCs).
Materials And Methods:
The binding of miR-21-5p to LncRNA MEG3 and SOD3 was determined using luciferase reporter assay; fluorescence quantitative PCR was used to detect the expression of LncRNA MEG3 at different induction times. hBMSCs were transfected with LncRNA MEG3 overexpression vector and induced for osteoblasts for 14 days. Alkaline phosphatase (ALP) staining, and alizarin red staining were used to detect bone differentiation, immunofluorescence assays were used to detect the expression of SOD3 and COL2A1.
Results:
Luciferase reporter assay revealed that miR-21-5p bond to LncRNA MEG3 and SOD3. Flow cytometry analysis showed that hBMSCs were highly pure. After osteogenic induction for 14 days, compared with the control group, the overexpression of LncRNA MEG3 significantly increased the activity of ALP and enhanced the formation of calcium nodules in hBMSCs. The overexpression also increased the expression of COL2A1 and SOD3 significantly (P<0.05).
Conclusions:
LncRNA MEG3 can promote the osteogenesis and bone regeneration of hBMSCs and increasing the expression of SOD3 and COL2A1 via targeting the miR-21-5p/SOD3 axis.
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