title: Different Methylation Patterns of RUNX2, OSX, DLX5 and BSP in Osteoblastic Differentiation of Mesenchymal Stem Cells.
abstract: OBJECTIVE
  Runt-related transcription factor 2 (RUNX2) and osterix (OSX) as two specific osteoblast transcription factors and distal-less homeobox 5 (DLX5) as a non-specific one are of paramount importance in regulating osteoblast related genes including osteocalcin, bone sialoprotein (BSP), osteopontin and collagen type Iα1. The present study sets out to investigate whether epigenetic regulation of these genes is important in osteoblastic differentiation of mesenchymal stem cells (MSCs).
  
  
  METHODS
  In this experimental study, MSCs were differentiated to osteoblasts under the influence of the osteogenic differentiation medium. DNA and RNA were extracted at days 0, 7, 14 and 21 from MSCs differentiating to osteoblasts. Promoter regions of RUNX2, OSX, DLX5 and BSP were analyzed by methylation-specific PCR (MSP). Gene expression was analyzed during osteoblastic differentiation by quantitative real-time polymerase chain reaction (PCR).
  
  
  RESULTS
  MSP analysis revealed that promoter methylation status did not change in RUNX2, DLX5 and BSP during MSC osteoblastic differentiation. In contrast, OSX promoter showed a dynamic change in methylation pattern. Moreover, RUNX2, OSX, DLX5 and BSP promoter regions showed three different methylation patterns during MSC differentiation. Gene expression analyses confirmed these results.
  
  
  CONCLUSIONS
  The results show that in differentiation of MSCs to osteoblasts, epigenetic regulation of OSX may play a leading role.
authors: Department of Hematology, Tarbiat Modares University, Tehran, Iran ; Hematology and Oncology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
authors: Majid
authors: M
authors: Farshdousti Hagh
authors: Department of Hematology, Tarbiat Modares University, Tehran, Iran ; Department of Medical Genetics, Tarbiat Modares University, Tehran, Iran ; Sarem Cell Research Center (SCRC), Sarem Women's Hospital, Tehran, Iran.
authors: Mehrdad
authors: M
authors: Noruzinia
authors: Department of Hematology, Zanjan University of Medical Science, Zanjan, Iran.
authors: Yousef
authors: Y
authors: Mortazavi
authors: Department of Hematology, Tarbiat Modares University, Tehran, Iran.
authors: Masood
authors: M
authors: Soleimani
authors: Department of Hematology, Tarbiat Modares University, Tehran, Iran.
authors: Saeed
authors: S
authors: Kaviani
authors: Department of Hematology, Tarbiat Modares University, Tehran, Iran.
authors: Saeed
authors: S
authors: Abroun
authors: Sarem Cell Research Center (SCRC), Sarem Women's Hospital, Tehran, Iran.
authors: Ali
authors: A
authors: Dehghani Fard
authors: Sarem Cell Research Center (SCRC), Sarem Women's Hospital, Tehran, Iran.
authors: Maryam
authors: M
authors: Mahmoodinia
keywords: DNA Methylation; Mesenchymal; Osteoblasts