[Study on LncRNA H19 promoting osteogenic differentiation of umbilical cord mesenchymal stem cells]

To observe the effects of long non-coding RNA H19(lncRNA H19) transfection on the expression of multiple osteogenic and angiogenic factors and the osteogenic capacity in human umbilical cord mesenchymal stem cells (hUC-MSCs), so as to provide a preliminary research basis for the future development of bone defect fillers with favorable blood supply.

Bioinformatics analysis was performed to confirm that H19 could regulate bone formation-related microRNAs (miRNAs) and their downstream osteogenic and angiogenic factors. The target molecules were finally identified as miR-137-3p, miR-383-5p, and their regulated factors including integrin-binding sialoprotein (IBSP), Runt-related transcription factor 2 (RUNX2), and fibroblast growth factor 5 (FGF5). Human umbilical cord mesenchymal stem cells were cultured in vitro. lncRNA H19 was then transfected into hUC-MSCs. Reverse transcription-polymerase chain reaction(RT-PCR) was used to detect the expression abundance of miR-137-3p and miR-383-5p in transfected cells. Western blot was applied to measure the expression levels of IBSP, Runx2 and FGF5. Alkaline phosphatase (ALP) staining and alizarin red S staining were conducted to evaluate the osteogenic capacity of cells after H19 transfection. We verified whether lncRNA H19 transfection could downregulate the expression of miR-137-3p and miR-383-5p, thereby upregulating the expression of osteogenic and angiogenic factors IBSP, Runx2 and FGF5, and confirmed that lncRNA H19 transfection promoted osteogenic differentiation of hUC-MSCs.

The expression levels of miR-137-3p and miR-383-5p were downregulated in hUC-MSCs transfected with lncRNA H19, whereas the expression levels of Runx2, IBSP and FGF5 were significantly increased from (0.533±0.058), (0.633±0.058), (0.500±0.100) before transfection to (1.300±1.100), (1.233±0.058), (0.967±0.115) after transfection, respectively, with statistically significant differences (P=0.000, P=0.000, P=0.006). Moreover, hUC-MSCs exhibited enhanced osteogenic capacity after H19 transfection.

Transfection with lncRNA H19 promotes the expression of several osteogenic and angiogenic factors and enhances the osteogenic capacity of hUC-MSCs. This provides a potential basis for the future development of tissue-engineered bone incorporating lncRNA H19 to improve its osteogenic and angiogenic performance.