Long non‐coding RNA‐H19 stimulates osteogenic differentiation of bone marrow mesenchymal stem cells via the microRNA‐149/SDF‐1 axis

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.

LncRNA H19 promotes odontoblastic differentiation of human dental pulp stem cells by regulating miR-140-5p and BMP-2/FGF9

“…Our results were consistent with Zeng et al [22]. H19 plays a critical role in the differentiation of many vital organs and plays pivotal roles in the differentiation of mesenchymal stem cells [23][24][25][26]. Keniry et al [27] found that H19 can enhance the expression level of RUNX2 to promote bone regeneration.…”

The lncRNA H19/miR-541-3p/Wnt/β-catenin axis plays a vital role in melatonin-mediated osteogenic differentiation of bone marrow mesenchymal stem cells

“…LncRNA NEAT1 promotes osteogenic differentiation of human BMSCs (hBMSCs) by modulating miR-29b-3p/BMP1 [ 21 ]. LncRNA H19 is a member of the lncRNA family, and studies have shown that up-regulated H19 facilitates osteogenic differentiation of BMSCs by facilitating stromal cell-derived factor 1 (SDF-1) through miR-149 [ 22 ]. Besides, H19 strengthens osteogenic differentiation of BMSCs by regulating the miR-140-5p/SATB2 axis [ 23 ].…”

The Involvement of Long Non-Coding RNAs in Bone

“…After establishing an in vitro rat-derived osteogenic differentiation model, Li et al [57] compared H19 expression between cells cultured in osteogenic condition compared with control, revealing its increased expression in differentiated cells with respect to cells without treatment. Additional experiments showed that H19 overexpression by H19 mimic treatment induces a relevant increase of ALP activity and calcium nodule formation, as well as an increase of osteogenic-related protein levels (i.e., ALP, OCN, RUNX2, and OSX), as opposed to cells transfected with shRNA.…”