Study On The Relationship Between Arhgap29 Gene Polymorphism And Non-syndromic Cleft Lip With Or Without Cleft Palate In Ningxia Population

Objective:Cleft lip and palate is the most common congenital cranial and maxillofacial dysplasia in human,the occurrence of this disease involves many factors.In recent years,finding and locating susceptibility genes has become a hotspot in the etiology of cleft lip and palate.This study is based on the previous research results of our project group,Arhgap29 gene was selected as the candidate gene for cleft lip and palate to explore the correlation between single nucleotide polymorphism of Arhgap29 gene and nonsyndrome cleft lip and palate in Ningxia.Methods:Blood samples were collected from 454 children with NSCL/P meeting the inclusion criteria(252 boys,202 girls),and 435 normal neonatal umbilical cord blood as control group(214 boys,221 girls),in the Department of Oral and Maxillofacial surgery,affiliated Hospital of Ningxia Medical University.5ml of venous blood from patients that met the inclusion criteria and cord blood from normal newborns were extracted respectively and then extracted the genomic DNA of them.The extracted DNA was identified for quality,including the concentration of DNA,purity identification and fragment length measurement etc.23 common SNP loci on Arhgap29,meet minor allele frequency(MAF)> 0.05,were selected in this study and respectively genotyped by Agri Seq TM targeting sequencing technique in the case group and the control group.The genotyping results were tested by Hardy-Weinberg equilibrium test,those SNPS met H-W equilibrium were analyzed by haploview for single-point correlation analysis,multi-spot linkage analysis and haplotype analysis.Results: In this study,the genotype distribution of 23 SNP loci by the HardyWeinberg equilibrium test showed no statistical difference in the control group.Comparing the case group with the control group,it was found that there were significant differences in the genotypic and allogenic frequencies of the four gene loci(rs3814019,rs72964308,rs7540684 and rs17396055)in the case group.Linkage analysis showed that a total of 23 SNPS on Arhgap29 gene formed 2 linkage blocks,and 2 haplotype of each block showed statistical differences between the case group and the control group(Block1: haplotype CGCAAGGGCGC(P=0.0406)and haplotype AATCGGGGCGT(P=0.0479);Block 2: haplotype CGTGCTCGGAGG(P=0.0103)and haplotype CGTATTCAAAGG(P=0.0336)).Conclusions:Among the 23 SNP sites of Arhgap29 gene,4 loci(rs3814019,rs72964308,rs7540684 and rs17396055)and some special haplotype combinations in the 2 blocks(CGCAAGGGCGC、AATCGGGGCGT、CGTGCTCGGAGG、CGTATTCAAAGG)were associated with the non-syndromic cleft lip with or without cleft palate among the population of Ningxia.