Association Study of Coronary Atherosclerotic Disease and Polymorphisms in TNFSF4 and TNFRSF4 Gene in a Chinese Han Population
|Keywords||Coronary atherosclerotic heart disease Single nucleotide polymorphisms Correlation Analysis TNFSF4 TNFRSF4 Involvement in patients with coronary artery branch|
[Background]: many studies have shown that coronary atherosclerotic heart disease is an inflammatory disease. During the development of inflammation, activated CD4 and CD8 T cells play an important role, and their activation requires a variety of signaling pathways stimulated; inter main stimulatory signal peptide and MHC-antigen B7-CD28 to form a double signal effect passage way, the auxiliary signal path T cell activation pathway is essential to the process conditions. Tumor necrosis factor superfamily member (TNFSF) and tumor necrosis factor receptor superfamily member (TNFRSF) T cell activation can be formed between the various necessary auxiliary signaling pathways, such as CD40/CD40L, OX40/OX40L ,4-1BB / 4-1BBL, CD70/CD27 and TLlA/DR3 so on. Through multiple populations association analysis and differential gene expression studies have shown that, OX40/OX40L signaling pathway in coronary atherosclerotic heart disease has an important role in the development process. Genetics and functional studies suggest that human TNFSF4 gene (encoding OX40L) and TNFRSF4 gene (encoding OX40) probably as important coronary atherosclerotic heart disease susceptibility genes play a role in the disease. Because gene polymorphisms associated with complex diseases with population specific, currently in Shandong Han population are not yet right TNFSF4 gene and TNFRSF4 gene polymorphisms and coronary atherosclerotic heart disease correlation studies. [Objective]: the use of case - control association study Method TNFSF4 gene within five single nucleotide polymorphisms (SNPs) and TNFRSF4 gene within a SNP and Shandong Han population coronary atherosclerotic heart disease The correlation between these polymorphic loci were: TNFSF4 gene promoter region 2 SNPsrs1234314 and rs45454293, the first intron region 2 tag SNPs rs3850641 and rsl234313, the second intron regions a tag SNP rs3861950; TNFRSF4 gene fifth intron 1 and functional sites in perfect linkage disequilibrium with rs2298211 SNP loci agent rs2298212. [Methods]: 1. Patient group and control group samples were collected: Shandong Han population collected in coronary atherosclerosis heart disease and normal controls, extracted blood and extracted genomic DNA. The collected coronary atherosclerotic heart disease patients met the diagnostic criteria for coronary artery disease, coronary angiography parallel identification; normal controls were Han and unrelated, at least within three generations living in healthy people of Shandong random individuals, and through history, physical examination, electrocardiogram and chest X-ray to exclude organic heart disease. 2 genotyping, genotype and allele frequency statistics: Polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP) method in the case and control groups in all the individuals of each polymorphic loci genotyping. According to results of agarose gel electrophoresis to determine individual genotypes, calculated at each locus in the case group and control group genotype and allele frequencies. 3. Hardy-Weinberg equilibrium goodness of fit test: According to the Hardy-Weinberg equilibrium calculation of polymorphic loci in each case and control groups in the expected genotype frequencies using the chi-square test to compare the observed genotype frequencies with the expected genotype differences between frequencies to P lt; 0.05 indicates a statistically significant difference. If P gt; 0.05 indicates the selected case and control groups in genetic equilibrium state, with group representation. 4 genotype and allele distribution between detection: the use of the chi-square test to compare each polymorphic loci in case and control groups between genotype and allele frequency differences to P lt; 0.05 represents the difference was statistically significance. If a polymorphic locus genotype and allele frequencies in a significant difference between the groups, namely P <0.05, indicating that the polymorphic loci with coronary atherosclerotic heart disease correlated; otherwise associated with this disease no correlation. In order to verify whether the distribution of genotypes and allele frequencies of gender specificity, especially for men and women stratified analysis, observed in different gender, each polymorphic loci with coronary atherosclerotic heart disease is correlated to P < 0.05 indicates a statistically significant difference. 5 cases of group involvement coronary bifurcation analysis: In accordance with coronary artery involvement count for the case group were stratified to Judkins method underwent selective coronary angiography, according to the number of diseased vessels angiography statistics (left main, left anterior descending artery, circumflex branch, right coronary artery or its major branch stenosis ≥ 50%), divided into 2, 3-vessel disease; involvement counts observed between different genotypes and allele frequencies whether differences. 6 effectiveness of existing statistical sample test: the use of PGA software (Power for Genetic Association Analysis, PGA) For existing samples can provide statistical power, in order to avoid errors caused by class Ⅱ false negative results. [Results] 1. TNFSF4 gene rs1234314, rs45454293, rs3850641, rs1234313, rs3861950 locus in Chinese Han population polymorphism; five points in the case group and the control group are in genetic equilibrium; genotypic and allelic distribution of gene frequencies between cases and control groups showed no significant difference between (P gt; 0.05). Comparison between different genotypes and involvement count allele frequencies found in the coronary involvement between you and each branch point difference in genotype and allele frequency distribution is also not statistically significant. 2. TNFRSF4 gene rs2298212 polymorphism loci in Han population in Shandong polymorphism; in the case group and the control group are in genetic equilibrium; genotype and allele frequencies between cases and control group was not statistically difference (P gt; 0.05). Comparison between different genotypes and involvement count allele frequencies found in the involvement of a coronary artery and three between genotype and allele frequency distributions that are of significant differences (P <0.05). 3 Application PGA software, test level α = 0.05, a rare allele frequency is assumed to be 0.17,0 R is assumed to be 1.4, linkage disequilibrium coefficient under the assumption of an existing statistical power of the sample exceeds 80%. [Conclusions] 1. TNFSF4 gene rs1234314, rs45454293, rs3850641, rs1234313, rs3861950 locus and Shandong Han population coronary atherosclerotic heart disease, no correlation between. The results need to be using the larger sample and different groups of case-control studies to verify. 2.OX40 gene rs2298212G / A polymorphic loci in Han population in Shandong coronary atherosclerotic disease severity correlated. The results need to be using the larger sample and different groups of case-control studies to verify.