Association Of The CYP2A6 Gene Polymorphisms With Tobacco Metabolin Related Coronary Heart Disease

OBJECTIVE:To explore four polymorphisms of CYP2A6(1,2,3 and 4)for association with tobacco metabolin related coronary heart disease;to explore a convenient method to optimize PCR conditions.METHODS:The genomic DNA was extracted from 205 patients with essential coronary heart disease(CHD)and 208 health adult Han subjects(NC)in Hunan.Based on their smoke habit,they were devided into smokers(SM)and nonsmokers groups(NS),207 and 206 subject respectively.Association analysis was employed.Uniform Design was applicated to optimize the four factors that may affect the polymerase chain reaction(RCR).The polymorphisms were genotyped by polymerase chain reaction-restriction fragment length polymorphism(RCR-RFLP).RESULTS:(1)The most favorable PCR conditions of CYP2A6 was: annealing temperature 61.2℃,DNA template content 50ng,the concentration of Mg²⁺1.5 mmol/L,Taq polymerase contents 1.SU.(2) The CYP2A6 genotypic frequency did not deviate from Hardy-Weinberg equilibrium in CHD and NC groups.There was no significantly difference of the distributions of four genotypes between the CHD and NC groups.(3)The CYP2A6 genotypic frequency did not deviate from Hardy-Weinberg equilibrium in SM and NS groups.CYP2A6^*1 frequency was higher in SM group than in NS group(SM:93.72%VS NS:87.86%,P＜0.05 X²=4.24).CYP2A6^*4 frequency was higher in NS group than in SM group(SM:5.31%VS NS:10.68%,P＜0.05 X²=4.04). Morbidity of CHD was higher in SM group than in NS group(SM: 62.93%VS NS:37.07%,X²=28.76,P＜0.01).CONCLUSION:(1)Uniform Design can act as a powerful tool for optimizing PCR reaction conditions.(2)CYP2A6 polymorphism may effect individual smoke habit.(3)Smoke may be a risk factor of coronary artery disease.(4)There was no correlationship between CYP2A6 polymorphisms and tobacco metabolin related cardiovascular disease.