Long non‚Äêcoding RNA‚ÄêH19 stimulates osteogenic differentiation of bone marrow mesenchymal stem cells via the microRNA‚Ä?49/ SDF‚Ä? axis
Authors: Shuanke Wang
Guangjie Li
Haiyan Zhao
Kaishan Ye
Xiangdong Yun
Xueliang Zhang
Yanhong Li
Jiangdong An
Xingwen Han
Journal: Journal of Cellular and Molecular Medicine (2020)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: bone defects; bone marrow mesenchymal stem cells; long non-coding RNA-H19; microRNA-149; osteogenic differentiation
Affiliations: Department of Orthopedics Lanzhou University Second Hospital Lanzhou China, The First Hospital of Lanzhou University Lanzhou China

Abstract:  Bone defects resulting from non-union fractures or tumour resections are common clinical problems. Long non-coding RNAs (lncRNAs) are reported to play vital roles in stem cell differentiation. The aim of this study was to elucidate the role of lncRNA-H19 in osteogenic differentiation of bone marrow mesenchymal stem cells (BMMSCs). Following the establishment of an osteogenic differentiation model in rats, the expression of H19, microRNA-149 (miR-149) and stromal cell-derived factor-1 (SDF-1) was measured by RT-qPCR. Thereafter, BMMSCs were isolated from rats and treated with a series of mimic, inhibitor or siRNA. SDF-1 expression, alkaline phosphatase (ALP) activity and osteocalcin (OCN) content were detected. The mineralized and calcified nodules were assessed by alizarin red S and Von Kossa staining. BMMSC surface markers were detected by flow cytometry. Western blot analysis was used to measure the expression of ALP, OCN, runt-related transcription factor 2 (RUNX2) and osterix (OSX) proteins. Lastly, dual-luciferase reporter gene assay and RNA immunoprecipitation were applied to verify the relationship of H19, miR-149 and SDF-1. Overexpressed H19 and SDF-1 and poorly expressed miR-149 were found in rats with osteogenic differentiation. H19 increased SDF-1 expression by binding to miR-149. H19 enhanced ALP activity, OCN content, calcium deposit and ALP, OCN, RUNX2 and OSX protein expression of BMMSCS by up-regulating SDF-1 via binding to miR-149. Taken together, up-regulated H19 could promote the osteogenic differentiation of BMMSCs by increasing SDF-1 via miR-149. 

Retracted: No

PubPeer comments:
 Figure 4C appears to have two images which overlap, after flipping horizontally and vertically. ***  , 
 Fig 3D.  Pearson s correlation analysis of the correlation between miR-149 and H19 . At right, my reconstruction, with best-fit trend line.  What is the origin of the diagonal line in 3D? It is far too steep to be a regression line. Fig 6B.  Pearson analysis of the correlation between miR-149 and SDF-1 . Again, the diagonal line seems arbitrary, with no relationship to the regression line.  , 
 Fig 1C.  Flow cytometry detection of the surface markers of BMMSCs (CD29, CD44, CD90 and CD45) .  I don t know how to make this comment without damaging Fig 1C s self-esteem, but flow-cytometry histograms shouldn t have overhangs. , 
 The original image concern seems to have been corrected, but the more serious concerns in #2 and #3 do not appear to have been addressed. *** Corrigendum. First published: 08 January 2022 In Guangjie Li, et al.,1 the H19 + mimic-NC image in Figure 4C contains error. The correct figure is shown below. The authors confirm all results, conclusions of this article remain unchanged.  , 
 #4 #3 I believe there may be several other examples of overhangs (added in pink, previously labelled in magenta). Do the authors have the original data used to generate these plots?  ,