Cui L, Johkura K, Takei S, Ogiwara N, Sasaki K. Structural differentiation, proliferation, and association of human embryonic stem cell-derived cardiomyocytes in vitro and in their extracardiac tissues. J Struct Biol 2007; 158(3): 307-17.
Zhang WJ, Park C, Arentson E, Choi K. Modulation of hematopoietic and endothelial cell differentiation from mouse embryonic stem cells by different culture conditions. Blood 2005; 105(1): 111-4.
Young JC, Varma A, DiGiusto D, Backer MP. Retention of quiescent hematopoietic cells with high proliferative potential during ex vivo stem cell culture. Blood 1996; 87(2): 545-56.
Tran NT, Trinh QM, Lee GM, Han YM. Efficient differentiation of human pluripotent stem cells into mesenchymal stem cells by modulating intracellular signaling pathways in a feeder/serum-free system. Stem Cells Dev 2012; 21(7): 1165-75.
Cardozo AJ, Gómez DE, Argibay PF. Transcriptional characterization of Wnt and Notch signaling pathways in neuronal differentiation of human adipose tissue-derived stem cells. J Mol Neurosci 2011; 44(3): 186-94.
Draper JS, Fox V. Human embryonic stem cells: multilineage differentiation and mechanisms of self-renewal. Arch Med Res 2003; 34(6): 558-64.
Siebert R, Willers CP, Opalka B. Role of the cyclin-dependent kinase 4 and 6 inhibitor gene family p15, p16, p18 and p19 in leukemia and lymphoma. Leuk Lymphoma 1996; 23(5-6): 505-20.
Chou BK, Mali P, Huang X, Ye Z, Dowey SN, Resar LM, et al. Efficient human iPS cell derivation by a non-integrating plasmid from blood cells with unique epigenetic and gene expression signatures. Cell Res 2011; 21(3): 518-29.
Bottardi S, Aumont A, Grosveld F, Milot E. Developmental stage-specific epigenetic control of human beta-globin gene expression is potentiated in hematopoietic progenitor cells prior to their transcriptional activation. Blood 2003; 102(12): 3989-97.
Holliday R. DNA methylation and epigenetic mechanisms. Cell Biophys 1989; 15(1-2): 15-20.
Zingg JM, Jones PA. Genetic and epigenetic aspects of DNA methylation on genome expression, evolution, mutation and carcinogenesis. Carcinogenesis 1997; 18(5): 869-82.
Sci J Iran Blood Transfus Organ 2013;10 (1): 31-39
Original Article
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
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