Ineffectiveness of Methylation in Rgulation of VHL, ECAD, and RUNX3 Genes in Erythroid Cells Differentiated by Erythropoietin
<h2>Background: Vast variety of intermediate factors including cell cycle regulators, growth factors, transcription factors, and signaling pathways are involved in hematopoietic stem cell (HSC) commitment and differentiation into distinct lineages. VHL, Ecad, and RUNX3 are among these. Epigene...
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| Format: | Article |
| Language: | English |
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Shahid Beheshti University of Medical Sciences
2015-04-01
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| Series: | Novelty in Biomedicine |
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| Online Access: | http://journals.sbmu.ac.ir/index.php/nbm/article/view/8676 |
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| author | Mehdi Azad Mehdi Goudarzi Ali Dehghanifard Mousa Vatanmakanian Mehdi Sahmani |
| author_facet | Mehdi Azad Mehdi Goudarzi Ali Dehghanifard Mousa Vatanmakanian Mehdi Sahmani |
| author_sort | Mehdi Azad |
| collection | DOAJ |
| description | <h2>Background: Vast variety of intermediate factors including cell cycle regulators, growth factors, transcription factors, and signaling pathways are involved in hematopoietic stem cell (HSC) commitment and differentiation into distinct lineages. VHL, Ecad, and RUNX3 are among these. Epigenetics is currently introduced as a potential mechanism to control the gene regulation. The aim of this study is to reveal the correlation between the expression level and methylation pattern of mentioned genes after in vitro differentiation of cord blood HSCs into erythroid lineage mediated by erythropoietin.</h2><p><strong>M</strong><strong>aterials and Methods: </strong>After isolation and expansion, the CD34+ cord blood stem cells were divided into two parts. The first part was used to extract the DNA and RNA and the second to differentiate into erythroid lineage. Methylation specific PCR (MSP) and Real-time PCR were used to determine the methylation status and expression levels of the genes, respectively.</p><p><strong>R</strong><strong>es</strong><strong>ults: </strong>Although the significant upregulation observed for VHL and Ecad genes and a down-regulation for RUNX3 gene after differentiation, no remarkable changes were seen in methylation pattern compared with cord blood HSCs by MSP technique.</p><p><strong>C</strong><strong>onclusion: </strong>It is appearing that methylation pattern in promoter region has not an effective role in expression of VHL, Ecad, and RUNX3. Moreover, considering the inability of MSP method to detect subtle differences in methylation level a more sensitive method is needed to distinguish the methylation levels of these genes before and after erythroid differentiation.</p> |
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| issn | 2345-3346 2345-3907 |
| language | English |
| last_indexed | 2024-04-13T13:37:25Z |
| publishDate | 2015-04-01 |
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| series | Novelty in Biomedicine |
| spelling | doaj.art-e97721e4fb2845029fc8a8730eb3aaf82022-12-22T02:44:44ZengShahid Beheshti University of Medical SciencesNovelty in Biomedicine2345-33462345-39072015-04-013257625030Ineffectiveness of Methylation in Rgulation of VHL, ECAD, and RUNX3 Genes in Erythroid Cells Differentiated by ErythropoietinMehdi Azad0Mehdi Goudarzi1Ali Dehghanifard2Mousa Vatanmakanian3Mehdi Sahmani4Department of Medical laboratory sciences, Faculty of Allied Medicine, Qazvin University of Medical Sciences, Qazvin, IranDepartment of Microbiology, School of Medicine, Shahid Beheshti University of Medical Science, Tehran, IranSarem Cell Research Center, Sarem Women’s Hospital, Tehran, IranDepartment of Hematology, Allied Medical School, Tehran University of Medical Sciences, Tehran, IranDepartment of Clinical Biochemistry and Genetics, Cellular and Molecular Research Centre, Qazvin University of Medical Sciences, Qazvin, Iran<h2>Background: Vast variety of intermediate factors including cell cycle regulators, growth factors, transcription factors, and signaling pathways are involved in hematopoietic stem cell (HSC) commitment and differentiation into distinct lineages. VHL, Ecad, and RUNX3 are among these. Epigenetics is currently introduced as a potential mechanism to control the gene regulation. The aim of this study is to reveal the correlation between the expression level and methylation pattern of mentioned genes after in vitro differentiation of cord blood HSCs into erythroid lineage mediated by erythropoietin.</h2><p><strong>M</strong><strong>aterials and Methods: </strong>After isolation and expansion, the CD34+ cord blood stem cells were divided into two parts. The first part was used to extract the DNA and RNA and the second to differentiate into erythroid lineage. Methylation specific PCR (MSP) and Real-time PCR were used to determine the methylation status and expression levels of the genes, respectively.</p><p><strong>R</strong><strong>es</strong><strong>ults: </strong>Although the significant upregulation observed for VHL and Ecad genes and a down-regulation for RUNX3 gene after differentiation, no remarkable changes were seen in methylation pattern compared with cord blood HSCs by MSP technique.</p><p><strong>C</strong><strong>onclusion: </strong>It is appearing that methylation pattern in promoter region has not an effective role in expression of VHL, Ecad, and RUNX3. Moreover, considering the inability of MSP method to detect subtle differences in methylation level a more sensitive method is needed to distinguish the methylation levels of these genes before and after erythroid differentiation.</p>http://journals.sbmu.ac.ir/index.php/nbm/article/view/8676Methylation, gene expression, erythropoietin, differentiation |
| spellingShingle | Mehdi Azad Mehdi Goudarzi Ali Dehghanifard Mousa Vatanmakanian Mehdi Sahmani Ineffectiveness of Methylation in Rgulation of VHL, ECAD, and RUNX3 Genes in Erythroid Cells Differentiated by Erythropoietin Novelty in Biomedicine Methylation, gene expression, erythropoietin, differentiation |
| title | Ineffectiveness of Methylation in Rgulation of VHL, ECAD, and RUNX3 Genes in Erythroid Cells Differentiated by Erythropoietin |
| title_full | Ineffectiveness of Methylation in Rgulation of VHL, ECAD, and RUNX3 Genes in Erythroid Cells Differentiated by Erythropoietin |
| title_fullStr | Ineffectiveness of Methylation in Rgulation of VHL, ECAD, and RUNX3 Genes in Erythroid Cells Differentiated by Erythropoietin |
| title_full_unstemmed | Ineffectiveness of Methylation in Rgulation of VHL, ECAD, and RUNX3 Genes in Erythroid Cells Differentiated by Erythropoietin |
| title_short | Ineffectiveness of Methylation in Rgulation of VHL, ECAD, and RUNX3 Genes in Erythroid Cells Differentiated by Erythropoietin |
| title_sort | ineffectiveness of methylation in rgulation of vhl ecad and runx3 genes in erythroid cells differentiated by erythropoietin |
| topic | Methylation, gene expression, erythropoietin, differentiation |
| url | http://journals.sbmu.ac.ir/index.php/nbm/article/view/8676 |
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