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Evaluation of methylation pattern in the promoter regions of P15 and P16 genes and the expression status of these factors in cord blood CD34+ stem cells
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M. Azad   , S. Kaviani , Y. Mortazavi , M. Norouzinia , M. Soleimani , S. Abroun , Z. Zonoubi , A. Atashi |
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| Keywords: Key words: Methylation, Stem Cells, Cord Blood |
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Type of Study: Research |
Subject:
Stem cells
Published: 2013/08/14
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Sci J Iran Blood Transfus Organ 2013;10 (1): 31-39
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Evaluation of methylation pattern in the promoter regions
of P15 and P16 genes and the expression status of these
factors in cord blood CD34+ stem cells
Azad M.1, Kaviani S.1, Mortazavi Y.2, Norouzinia M. 1, Soleimani M.1, Abroun S.1,
Zonoubi Z.3, Atashi A.1
1Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
2Zanjan University of Medical Sciences, Zanjan, Iran
3Faculty of Medical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Abstract
Background and Objectives
Cell signaling elements such as P15 and P16 play an important role in differentiation of primitive cells into various cell types. These factors are involved in controlling gene expression by different mechanisms, of which epigenetics especially methylation is noteworthy. The main objectives of this study were discovering the expression status of target genes in CD34 + cord blood stem cells and determining the methylation changes in the respective genes is this same stage.
Materials and Methods
After collection of cord blood bags and purification and proliferation of stem cells, cellular DNA was isolated. In later stages, cDNA and Bisulfite treated DNAs were synthesized from RNA and DNA of primary cells, respectively. Polymerase Chain Reaction and Methylation Specific PCR were later performed for both genes.
Results
After MSP, it was found that the P15 has partial methylation and expression in CD34 + stem cells, and P16 lacks methylation in this stage and is completely expressed. The results of PCR indicated expression of both genes in CD34 + stem cells.
Conclusions
Gene expression profile of each tissue or cell is in accordance with its functions; the specific expression of P15 and P16 can indicate a possible role of these genes in biology of CD34 + stem cells in umbilical cord blood. There is always substantial agreement between expression of a gene and its epigenetic changes.
Key words: Methylation, Stem Cells, Cord Blood
Received: 28 Mar 2012
Accepted:18 Jul 2012
Correspondence: Kaviani S., PhD of Hematology. Associate Professor of Faculty of Medical Sciences, Tarbiat Modares University.
P.O.Box: 14115-331, Tehran, Iran. Tel: (+9821) 82883832; Fax: (+9821) 82884507
E-mail:
kavianis@modares.ac.ir
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Azad M, Kaviani S, Mortazavi Y, Norouzinia M, Soleimani M, Abroun S, et al . Evaluation of methylation pattern in the promoter regions of P15 and P16 genes and the expression status of these factors in cord blood CD34+ stem cells . Sci J Iran Blood Transfus Organ 2013; 10 (1) :31-39 URL: http://bloodjournal.ir/article-1-737-en.html
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