Direct Bisulfite Sequencing and Methylation Specific PCR to Detect Methylation of p15INK4b and F7 genes in Coronary Artery Disease Patients
Genome-Wide Association Studies (GWAS) have identified genetic variants contributing to the risk of cardiovascular disease (CVD) at the chromosome 9p21 locus. The chromosome 9p21 is an important susceptibility locus for several multifactorial diseases like ischemic stroke, aortic aneurysm, type 2 di...
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University of Tehran
2016-01-01
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| Series: | Journal of Sciences, Islamic Republic of Iran |
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| Online Access: | https://jsciences.ut.ac.ir/article_56005_e24ebf02e8a108a49701ab41119c5dac.pdf |
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| author | M. Yari A. Movafagh A. Sayad M. D. Omrani M. A. Broumand K. Majidzadeh-A R. Mirfakhraie A. Khoshdel K. Amini |
| author_facet | M. Yari A. Movafagh A. Sayad M. D. Omrani M. A. Broumand K. Majidzadeh-A R. Mirfakhraie A. Khoshdel K. Amini |
| author_sort | M. Yari |
| collection | DOAJ |
| description | Genome-Wide Association Studies (GWAS) have identified genetic variants contributing to the risk of cardiovascular disease (CVD) at the chromosome 9p21 locus. The chromosome 9p21 is an important susceptibility locus for several multifactorial diseases like ischemic stroke, aortic aneurysm, type 2 diabetes mellitus and coronary artery disease (CAD). F7 gene because of its role in activating the extrinsic pathway by the exposure of tissue factor after plaque disruption is related to atherothrombosis. The aim of the present study was to evaluate methylation status of two CAD related genes, p15INK4b and F7, in Iranian patients with coronary artery disease (CAD). Thirty samples from 15 male and 15 female with diagnosed 3 vessels disease CAD and 60 Samples from 60 non-CAD controls who underwent coronary angiography was analyzed by MSP and direct sequencing. DNA methylation levels at p15INK4b gene increased significantly in CAD patients in comparison with control group (p-value<0.001). To quantitative analysis of methylation, direct bisulfite sequencing method revealed 6 methylated CpGs islands. No significant difference in F7 promoter methylation was observed between CAD patients and control group. Based on the findings of this study it is possible to assume, p15INK4b methylation is associated with pathophysiology of CAD. |
| format | Article |
| id | doaj-art-5eb2d5e0b6d043a3adf7f28da8e7f321 |
| institution | Kabale University |
| issn | 1016-1104 2345-6914 |
| language | English |
| publishDate | 2016-01-01 |
| publisher | University of Tehran |
| record_format | Article |
| series | Journal of Sciences, Islamic Republic of Iran |
| spelling | doaj-art-5eb2d5e0b6d043a3adf7f28da8e7f3212025-08-20T03:53:47ZengUniversity of TehranJournal of Sciences, Islamic Republic of Iran1016-11042345-69142016-01-01271232956005Direct Bisulfite Sequencing and Methylation Specific PCR to Detect Methylation of p15INK4b and F7 genes in Coronary Artery Disease PatientsM. Yari0A. Movafagh1A. Sayad2M. D. Omrani3M. A. Broumand4K. Majidzadeh-A5R. Mirfakhraie6A. Khoshdel7K. Amini81 Department of Medical Genetics, Faculty of Medicine, University of Shahid Beheshti Medical Sciences, Tehran, Islamic Republic of Iran.1 Department of Medical Genetics, Faculty of Medicine, University of Shahid Beheshti Medical Sciences, Tehran, Islamic Republic of Iran.1 Department of Medical Genetics, Faculty of Medicine, University of Shahid Beheshti Medical Sciences, Tehran, Islamic Republic of Iran.1 Department of Medical Genetics, Faculty of Medicine, University of Shahid Beheshti Medical Sciences, Tehran, Islamic Republic of Iran.2 Department of Molecular Pathology, Tehran Heart Center, University of Tehran Medical Sciences, Tehran, Islamic Republic of Iran.3 Department of Breast Cancer Genetics, Breast Cancer Research Center (BCRC), University of Tehran, Tehran, Islamic Republic of Iran.1 Department of Medical Genetics, Faculty of Medicine, University of Shahid Beheshti Medical Sciences, Tehran, Islamic Republic of Iran.4 Department of Medical Sciences, Faculty of Medicine, University of AJA, Tehran, Islamic Republic of Iran.4 Department of Medical Sciences, Faculty of Medicine, University of AJA, Tehran, Islamic Republic of Iran.Genome-Wide Association Studies (GWAS) have identified genetic variants contributing to the risk of cardiovascular disease (CVD) at the chromosome 9p21 locus. The chromosome 9p21 is an important susceptibility locus for several multifactorial diseases like ischemic stroke, aortic aneurysm, type 2 diabetes mellitus and coronary artery disease (CAD). F7 gene because of its role in activating the extrinsic pathway by the exposure of tissue factor after plaque disruption is related to atherothrombosis. The aim of the present study was to evaluate methylation status of two CAD related genes, p15INK4b and F7, in Iranian patients with coronary artery disease (CAD). Thirty samples from 15 male and 15 female with diagnosed 3 vessels disease CAD and 60 Samples from 60 non-CAD controls who underwent coronary angiography was analyzed by MSP and direct sequencing. DNA methylation levels at p15INK4b gene increased significantly in CAD patients in comparison with control group (p-value<0.001). To quantitative analysis of methylation, direct bisulfite sequencing method revealed 6 methylated CpGs islands. No significant difference in F7 promoter methylation was observed between CAD patients and control group. Based on the findings of this study it is possible to assume, p15INK4b methylation is associated with pathophysiology of CAD.https://jsciences.ut.ac.ir/article_56005_e24ebf02e8a108a49701ab41119c5dac.pdfcoronary artery disease (cad)chromosome 9p21p15ink4b genef7 geneepigenetics |
| spellingShingle | M. Yari A. Movafagh A. Sayad M. D. Omrani M. A. Broumand K. Majidzadeh-A R. Mirfakhraie A. Khoshdel K. Amini Direct Bisulfite Sequencing and Methylation Specific PCR to Detect Methylation of p15INK4b and F7 genes in Coronary Artery Disease Patients Journal of Sciences, Islamic Republic of Iran coronary artery disease (cad) chromosome 9p21 p15ink4b gene f7 gene epigenetics |
| title | Direct Bisulfite Sequencing and Methylation Specific PCR to Detect Methylation of p15INK4b and F7 genes in Coronary Artery Disease Patients |
| title_full | Direct Bisulfite Sequencing and Methylation Specific PCR to Detect Methylation of p15INK4b and F7 genes in Coronary Artery Disease Patients |
| title_fullStr | Direct Bisulfite Sequencing and Methylation Specific PCR to Detect Methylation of p15INK4b and F7 genes in Coronary Artery Disease Patients |
| title_full_unstemmed | Direct Bisulfite Sequencing and Methylation Specific PCR to Detect Methylation of p15INK4b and F7 genes in Coronary Artery Disease Patients |
| title_short | Direct Bisulfite Sequencing and Methylation Specific PCR to Detect Methylation of p15INK4b and F7 genes in Coronary Artery Disease Patients |
| title_sort | direct bisulfite sequencing and methylation specific pcr to detect methylation of p15ink4b and f7 genes in coronary artery disease patients |
| topic | coronary artery disease (cad) chromosome 9p21 p15ink4b gene f7 gene epigenetics |
| url | https://jsciences.ut.ac.ir/article_56005_e24ebf02e8a108a49701ab41119c5dac.pdf |
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