LncRNA H19 Targets miR-19b-3p/SOX9 Pathway to Promote Osteogenic Differentiation of Adipose-Derived Mesenchymal Stem Cells
Objective:To investigate the effects of LncRNA H19 targeting miR-19b-3p/SOX9 pathway on the osteogenic differentiation of adipose-derived mesenchymal stem cells(ADMSCs).Methods:Human ADMSCs were cultured and identified in vitro.The cells were divided into five groups:control group(regular culture),H19 group(transfected with LncRNA H19 lentivirus overexpression vector),LncRNA-NC group(transfected with lentivirus empty vector),miR-19b-3p group(co-transfected with LncRNA H19 lentivirus overexpression vector and miR-19b-3p mimic lentivirus vector),and miR-NC group(co-transfected with LncRNA H19 lentivirus overexpression vector and miR-NC lentivirus vector).The expression of LncRNA H19,miR-19b-3p,SOX9,zinc finger osteoblast-specific transcription factor(Osx),osteocalcin(OCN),and RUNX2 was detected in each group.ALP staining and Alizarin Red staining were observed in each group,cell proliferation activity and ALP activity were compared,and the expression of SOX9,BMP-2,and OPN proteins in each group was analyzed to verify the targeted relationship between LncRNA H19 and miR-19b-3p.Results:Compared with the control group and LncRNA-NC group,the expression of LncRNA H19 and SOX9,Osx,OCN,and RUNX2 mRNA,as well as the expression of SOX9,BMP-2,and OPN proteins,ALP activity,and the amount of mineralized nodule formation were increased in the H19 group,while the ex-pression of miR-19b-3p was decreased(P＜0.05).Compared with the H19 group and miR-NC group,the expression of LncRNA H19 and SOX9,Osx,OCN,and RUNX2 mRNA,as well as the expression of SOX9,BMP-2,and OPN proteins,ALP activity,and the amount of mineralized nodule formation were decreased in the miR-19b-3p group,while the expression of miR-19b-3p was increased(P＜0.05).There was no statis-tical difference in the absorbance of each group at 24h,48h,and 72h(P＞0.05).There was a targeted rela-tionship between LncRNA H19 and miR-19b-3p.Conclusion:LncRNA H19 can negatively regulate miR-19b-3p,upregulate SOX9,and contribute to the differentiation of human ADMSC cells into osteoblasts.Ln-cRNA H19 can negatively regulate miR-19b-3p,upregulate SOX9,and promote the osteogenic differentia-tion of human ADMSCs.
Long non coding ribonucleic acid H19MiR-19b-3p/SRY-box transcription factor 9 pathwayAdipose derived mesenchymal stem cellsOsteoblast differentiation
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