The Study On The Correlation Between Copy Number Variation Of KIAA1267, PRH1Gene And Congenital Heart Disease

Objective:Tested the incidence rate of KIAA1267(KAT8regulatory NSL complex subunit1) and PRH1(proline-rich protein HaeⅢ subfamily1) gene copy number variation (CNV) in the population and investigated its correlation with congenital heart disease.Method:The research is based on330cases of distributed congenital heart disease (CHD) patients, one case of CHD pedigrees and300cases of normal control samples, by using real-time polymerase chain reaction (Real-time PCR) technology to test the candidate genes KIAA1267and PRH1which were filtered out by array-based comparative genomic hybridization (aCGH) in the pre-experiment, detect their copy number variation and incidence in the population, and analyze statistically.Result:1. The results of the tested KIAA1267gene of the samples of sporadic congenital heart disease patients and statistical analysis:(1)330cases of congenital heart disease patients which are tested by the primers of KIAA1267-1and KIAA1267-2and the validation of three wells, reveals that8cases of samples have fragment deletions in the KIAA1267gene region, and322cases of samples have no exception.300cases of tested normal control samples have no fragment deletions in the KIAA1267gene region.(2)By using the primers KIAA1267-3and KIAA1267-6to locate the boundary to the8cases of positive samples which are proved of fragment deletions, it finds that the8cases of samples have fragment deletions with unequal size. The samples which are numbered as10HD173-1and10HD318-1have multiple copies of the variation near the area of the primer KIAA1267-2, that is deletion and the concurrency of amplification.(3)The copy number variation rate of KIAA1267gene of experimental group of congenital heart disease is greater than the normal control group (X2=0.008, P<0.01). 2. The tested about PHI gene of sporadic congenital heart disease samples and statistical analysis results:(1)In200cases of patients with congenital heart disease (amplified fragment located in the PRH1dean upstream) we found that zero copy in59cases (29.50%), one copy in23cases (11.50%), two copies in98cases (49.00%), many copies in20cases (10.00%) In normal control group samples of120cases we detected zero copy in14cases (11.67%), one copy in23cases (19.17%), two copies in77cases (64.17%), many copies in6cases (5.00%). According to the copy number, test results of the two groups of samples are divided into four grades, and then used of statistical software SPSS17.0ranked data of two independent samples comparison the Wilcox on rank-sum test analysis, obtained for Z=-4.386, P<0.01, significant differences, that is, PRH1gene copy number variation rate of congenital heart disease in experimental group is higher than the normal control group.(2) Though the validation of three wells and the verification of the primers of gene upstream sequence, the experimental results above are the same.(3) We detected the positive samples which are from the experimental results with PRH1-5primers (amplified fragment located in the PRH1).We found that all were2copies.3. The pedigree of congenital heart disease gene copy number variation detection:(1)Four people of the pedigree(Ⅰ1, Ⅰ2,Ⅱ2,Ⅱ3) were not found deletions in the KIAA1267gene.(2) Ⅰ2and Ⅱ2samples of genomic DNA in PRH1-2primer experiments without amplification, Ⅰ1and Ⅱ3were single copy, but all four samples were2copies in PRH1-5primer experiments.Conclusion:1. In congenital heart disease group we detected8cases that have copy number variation (deletion) in KIAA1267gene, two cases were deletion and amplification coexist; In the control group were not detected in deletion or amplification variation (X2=0.008, P <0.01), we consider that copy number variation in KIAA1267gene may be the risk factor of congenital heart disease.2. Through the PRH1gene detection and the Wilcox on rank-sum test statistical analysis (Z=-4.386, P<0.01), it suggested significant differences between the experimental results of two groups of samples. It considered that PRH1gene copy number variation may increase the risk of congenital heart disease.3. Targeting the situation that the deletion only be found in the PRH1dean upstream, but no deletion in PRH1.We considered that the pathogenic mechanisms may be related to the effects of gene expression regulation process, but the exact correlation remains to be further studied.