Abstract

To explore the influence of the long-non coding RNA-H19/microRNA-140-5p/Wnt1 axis on the osteogenic differentiation in bone marrow mesenchymal stem cells. First, we used dual-luciferase reporter gene detection to verify long-non coding-H19/miRNA-140-5p/Wnt1 regulatory axis targeting relationship. Then long-non coding-H19 was overexpressed or silenced in bone marrow mesenchymal stem cells and then long-non coding-H19, microRNA-140-5p and Wnt1 expression were assessed using quantitative real-time polymerase chain reaction and cell activity was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide. Then the osteogenic differentiation culture was performed, formation of calcium nodules were assessed via Alizarin red staining, while the expression of osteocalcin and runt-related transcription factor 2 detected via Western blot. Wild type long-non coding RNA-H19 group and wild type-Wnt1 group showed significantly decreased luciferase activity in dual-luciferase reporter assays (p<0.05). After overexpressed long-non coding RNA-H19, the microRNA-140-5p expression decreased and Wnt1 expression increased (p<0.05), and while silenced the long-non coding RNA-H19 expression, the opposite result was found (p<0.05). In terms of cell viability, there were no significant differences between groups after long-non coding RNA-H19 was overexpressed or silenced (p>0.05). After overexpressed long-non coding RNA-H19, calcium nodules formation, mineralized bone matrix level, the expression of osteocalcin and runt-related transcription factor 2 protein significantly increased (p<0.05), and while silenced the long-non coding RNA-H19 expression, the opposite result was found (p<0.05). Long-non coding RNA-H19 overexpression promotes the bone marrow mesenchymal stem cells osteogenic differentiation via targeting microRNA-140-5p/Wnt1 signaling pathway.