间充质干细胞成骨分化过程中RUNX2、OSX、DLX5和BSP的不同甲基化模式
Different Methylation Patterns of RUNX2, OSX, DLX5 and BSP in Osteoblastic Differentiation of Mesenchymal Stem Cells.
作者信息
Farshdousti Hagh Majid, Noruzinia Mehrdad, Mortazavi Yousef, Soleimani Masood, Kaviani Saeed, Abroun Saeed, Dehghani Fard Ali, Mahmoodinia Maryam
机构信息
Department of Hematology, Tarbiat Modares University, Tehran, Iran ; Hematology and Oncology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
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.
出版信息
Cell J. 2015 Spring;17(1):71-82. doi: 10.22074/cellj.2015.513. Epub 2015 Apr 8.
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).
MATERIALS AND 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.
CONCLUSION
The results show that in differentiation of MSCs to osteoblasts, epigenetic regulation of OSX may play a leading role.
目的
runt相关转录因子2（RUNX2）和osterix（OSX）作为两种特定的成骨细胞转录因子，而远端缺失同源盒5（DLX5）作为一种非特异性转录因子，在调节包括骨钙素、骨涎蛋白（BSP）、骨桥蛋白和Iα1型胶原蛋白在内的成骨细胞相关基因方面至关重要。本研究旨在探讨这些基因的表观遗传调控在间充质干细胞（MSC）向成骨细胞分化过程中是否重要。
材料与方法
在本实验研究中，MSC在成骨分化培养基的影响下分化为成骨细胞。在第0、7、14和21天从向成骨细胞分化的MSC中提取DNA和RNA。通过甲基化特异性PCR（MSP）分析RUNX2、OSX、DLX5和BSP的启动子区域。在成骨细胞分化过程中通过定量实时聚合酶链反应（PCR）分析基因表达。
结果
MSP分析显示，在MSC向成骨细胞分化过程中，RUNX2、DLX5和BSP的启动子甲基化状态没有变化。相比之下，OSX启动子的甲基化模式呈现动态变化。此外，在MSC分化过程中，RUNX2、OSX、DLX5和BSP启动子区域呈现三种不同的甲基化模式。基因表达分析证实了这些结果。
结论
结果表明，在MSC向成骨细胞分化过程中，OSX的表观遗传调控可能起主导作用。
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