Analysis of Differential Translation Profiles of Human Bone Marrow Mesenchymal Stem Cells during Osteogenic Differentiation.

Abstract

Objective: To explore the differential translation profiles and coding products of human jaw bone marrow mesenchymal stem cells (h-JBMMSCs) during osteogenic differentiation.

Methods: Ribo-seq was used to examine the differential translated genes (DEGs), open reading frames (ORFs) and genes associated with the osteogenic differentiation phase of h-JBMMSCs. Western blotting (WB) was performed to detect the expression of osteocalcin (OCN) and bone sialoprotein (BSP). Alkaline phosphatase (ALP) activity and alizarin red staining (ARS) were used to detect osteogenic differentiation. A lentivirus containing 5'UTR-ORF-GFPmut was designed to transfect h-JBMMSCs, and fluorescence and green fluorescent protein (GFP) expression were analysed. The SNHG1 peptide was synthesised for osteogenic induction and to detect osteogenic markers.

Results: A total of 53,432 ORFs were detected and 199 candidate translation sORFs, including lncRNA SNHG1, were identified after removing the annotated protein-coding genes. In addition, the 5'UTR-ORF-GFPmut showed green fluorescence and expressed GFP. Knockdown of the lncRNA SNHG1 increased the ALP activity of h-JBMMSCs, promoted the expression of OCN and BSP, and enhanced the intensity of ARS and calcium ion content. However, overexpression of lncRNA SNHG1 and the SNHG1 polypeptide inhibited the osteogenic differentiation of h-JBMMSCs.

Conclusion: LncRNA SNHG1 inhibited the osteogenic differentiation of h-JBMMSCs. LncRNA SNHG1 can encode a peptide of 19-amino acid and inhibit the osteogenic differentiation of h-JBMMSCs.