Research Of The Relationship Between TBX5 Gene And Congenital Heart Disease

ObjectiveA case controlled study using the patients with congenital heart disease as the subjects and normal persons as the controls.To explore the association between single nucleotide polymorphism of TBX5 and congenital heart disease,high resolution melting curve(HRM)was used to detect single nucleotide polymorphism of TBX5 in non-coding region.We explore the effects of interaction between environmental exposure factors and genetic polymorphism in TBX5 and MTHFR on risk of congenital heart disease.This study was to comprehensively understand and explain the pathogeny of congenital heart disease.Then we could provide scientific basis of identification of high-risk population and prevention and control of congenital heart disease.Methods1.A case-control study of 186 patients with congenital heart disease and 200 healthy children in Qilu Children's Hospital of Shandong University was performed exposure to analyse some exposure factors in environment that related with congenital heart disease.2.Molecular biology technique was used to extract DNA.The non-coding region of TBX5 gene was amplified by PCR.Single nucleotide polymorphism of TBX5 was analyzed by high resolution melting curve and then validated by gene sequencing.3.Statistical analysis was performed using the Statistical Package for Social Sciences(SPSS 22.0 for Windows;IBM).Differences between groups were analyzed by the Student t test for continuous variables.Differences incategorical variables,genotype distribution,and the Hardy-Weinberg equilibrium were tested by?2analysis.The SNP�SNP interaction and the SNP�environment interaction was determined by multifactor dimensionality reduction(MDR).Results1.Single nucleotide polymorphism of TBX5 was successfully detected by high resolution melting curve.2.The genotype frequencies were in agreement with those predicted by the Hardy-Weinsberg equilibrium(?~2=0.378,P=0.539).3.The allele mutation frequencies of TBX5 in the control group and case group were42.50%and 50.27%respectively,the differences were significant between the two groups(?~2=4.68,P=0.031).The allele frequencies of TBX5 rs 883079(G/G,G/A,A/A)for the control group were 32.00%,51.00%and 17.00%.The allele frequencies in patients were27.96%,43.55%and 28.49%,the differences were significant between the two groups(?~2=4.47,P=0.035).Comparing with G/G genotype in TBX5 gene rs 883079,A/A genotype increased the risk of congenital heart disease with OR=1.919 and 95%CI:1.090-3.376.Comparing with G allele,A allele may be an independent risk factor for congenital heart disease in the Chinese Han population(OR=1.368,95%CI:1.030-1.817).4.Univariate logistic regression analysis showed the risk factors of congenital heart disease:childbearing ages(OR=1.123,95%CI:1.025-1.23),pregnancy induced hyperten-sion(OR=3.174,95%CI:1.000-10.077),history of abortion(OR=2.414,95%CI:1.122-5.194),history of taking contraceptive(OR=1.870,95%CI:1.006-3.475),history of having a cold(OR=1.744,95%CI:1.067-2.852),and mental stimulation(OR=1.940,95%CI:1.092-3.447)and emotion(OR=0.701,95%CI:0.544-0.904).Multivariate logistic regression analysis showed that pregnancy induced hypertension was an increased risk of congenital heart disease(OR=3.824,95%CI:1.080-13.542),emotion may be an independent protective factor for congenital heart disease(OR=0.664,95%CI:0.506-0.873).5.The optimal model containing emotion,MTHFR C677T(rs 1801133),TBX5 gene rs 883079 obtained with multifactor dimensionality reduction was found to be statistically significant.The Cross-Validation consistency was highest.Conclusions1.Statistically significant association was found between single nucleotide polymor-phism of TBX5(rs 883079)and congenital heart disease.Comparing with G/G genotype in TBX5 gene rs 883079,A/A genotype increased the risk of congenital heart disease,may be an independent risk factor for congenital heart disease(OR=1.919,95%CI:1.090-3.376).A allele may be an independent risk factor for congenital heart disease(OR=1.368,95%CI:1.030-1.817).2.Pregnancy induced hypertension were an increased risk of congenital heart disease(OR=3.174,95%CI:1.000-10.077),emotion may be an independent protective factor for congenital heart disease(OR=0.664,95%CI:0.506-0.873).3.The genetic polymorphisms of gene might contribute to congenital heart disease susceptibility and interact with environmental exposure by multifactor dimensionality red-uction.There are Interaction between TBX5 gene rs 883079,MTHFR C677T(rs 1801133)and emotion factors on congential heart disease.