| Coronary artery disease (CAD) is a polygenic disorder, resulting from a variety of factors and diseases. Many risk factors are associated with development of CAD which is considered as an important disease that controlled by epigenetic. DNA methylation is a critical epigenetic modification that silences gene transcription, without changing DNA sequence. It has been found that DNA methylation plays a key role in initiation and progress of cardiovascular disease. Acute coronary syndrome (ACS), including unstable angina (UA), acute myocardial infarction (AMI) and other acute myocardial ischemia are caused by decreased blood flow in the coronary arteries suddenly, and ACS has a high mortality. Thus, it is worth that to explore the further mechanism of DNA methylation participate in the development of ACS.Objectives: (1)Illumina’s Infinium human Methylation 450 Beadchip arrays were used to examine genome-wide DNA methylation profiles in 3 sample pairs from ACS group and the normal control group.(2) Using MSP to compare the methylation level of NCP1 promoter between CAD and control group. To provide experimental evidence for specific gene methylation level as new targets of coronary heart disease diagnosis and treatment.Methods:(1)Whole blood samples of 3 ACS paients and 3 normal persons were collected respectively for DNA extraction and bisulfite conversion. Illumina HD 450K Infinium Methylation BeadChip was applied to detect more than 450,000 methylation sites of the whole human genome, covering 96% CpG islands. For the probe methylation sites in Illumina HD 450K Infinium Methylation BeadChip, the data is then standardized preprocessing, and finally, calculate of the degree of methylation levels (Beta score). The hypermethylation genes were then analyzed by Gene Ontology (GO) and Kyoto encyclopedia of Genes and Genomes (KEGG) Analysis. (2)MSP was used to exam the methylation levels of NPC1 in the whole blood samples of 64 healthy subjects (control group),37 SCAD patients and 55 ACS patients.Results:(1)A total of 770 genes significant differed in methylation level between ACS patients and normal persons (Beta score>0.2, p<0.05), with 640 hypomethylation genes and 130 hypermethylation genes. The results of GO and KEGG suggest that the hypermethylated genes are associated with some biological processes, such as signaling pathway, glucose and lipid metabolism, cell adhesion molecules and apoptosis. The hypermethylated genes may one of the causes of ACS. (2)The methylation level of NPC1 promoter in SCAD patients is much higher than that in controls and ACS patients, while there are no difference between controls and ACS patients.Conclusions:(1)The analyzation of Illumina’s Infinium human Methylation 450 Beadchip arrays showed that:the methylation levels are significantly different between ACS group and normal group. This difference in the level of DNA methylation may be an important factor leading to the development of ACS, and laid the foundation for further explore the molecular mechanism of the ACS and new prevention methods. (2) The methylation level of NPC1 were significantly increased in patients with coronary heart disease. It is speculated that the NPC1 hypermethylation may accelerate the formation of plaques in the early stage of AS and stable the plaques in the late stage. To detect the methylation level of these genes would be benefit for earyl diagnosis and disease monitoring forcoronary artery disease patients, in particular ACS. |
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