Evaluation of expression and epigenetic changes of DLX5 and DLX6 genes in osteoblastic differentiation of bone marrow mesenchymal stem cell

Abstract

. In this study, the methylation status of the promoter region of DLX5 and DLX6 genes and their role in controlling osteoblastic differentiation in MSCs was investigated during the osteoblastic differentiation of MSCs. Methods: Mesenchymal stem cells were cultured under standard conditions and then differentiated into osteoblast lineage using differentiation medium. For this purpose, DMEM medium (10% FBS) containing L-glutamine, dexamethasone, beta-glycerol phosphate and ascorbate 2-phosphate medium were added within 21 days. Gene expression and promoter methylation of DLX5 and DLX6 genes were evaluated on days 7, 14 and 21 of osteoblast differentiation using Real time PCR and methylation specific-quantitative PCR (MS-QPCR), respectively. Results: DLX5 and DLX6 gene expression was up-regulated and promoter methylation decreased. 5-azacytidine caused a significant reduction in methylation and increased mRNA expression of DLX5 and DLX6.