COL6A2基因3′-UTR区域功能单核苷酸多态性位点rs1044598参与miR-4252调节中国汉族人群先天性房间隔缺损

The mechanism of mi R-4252 regulating pathogenesis of congenital atrial septal defect with SNP(rs1044598) in 3′-UTR of COL6A2 in Han Chinese populations

目的 :探讨COL6A2基因3′非翻译区(3′-UTR)单核苷酸多态性(single nucleotide polymorphism,SNP)位点对先天性房间隔缺损(congenital atrial septal defect,ASD)发病风险的影响及其可能的作用机制。方法:搜索Pub Med及Hapmap数据库获得COL6A2基因3′-UTR区域中国汉族人群最小等位基因频率>0.05的SNP位点,随后通过mi RNA-SNP功能网站预测SNP位点的功能情况并与本课题组前期ASD全基因组关联研究数据库比对,研究SNP位点与ASD的发病关联,最后对可能的功能位点进行功能学研究。结果:rs1044598位点AA基因型较野生TT基因型显著减少了36%的患病风险。HEK293T细胞、H9C2细胞以及SD乳鼠原代心肌细胞荧光素酶报告基因转染实验证实,mi R-4252与COL6A2基因rs1044598不同基因型表达质粒共转染后,荧光强度在3个细胞系中均有显著差异(P<0.05)。结论:COL6A2基因rs1044598位点的变异可能与ASD的发病风险相关。mi R-4252可通过与COL6A2的3′-UTR区域发生有效结合而下调基因的表达,而rs1044598位点的变异参与这一机制并减少ASD的发生。

Objective：To investigate the association and potential mechanism between single nucleotide polymorphism（SNP）in 3′untranslated region（3′-UTR） of COL6A2 and the risk of congenital atrial septal defect（ASD）. Methods：SNPs in 3′-UTR of COL6A2 were searched in Pub Med and Hapmap database for minimum allele frequency（MAF） 〉0.05 in Han Chinese population. Then the potential functional SNPs were predicted in website of mi RNA-SNP. Meanwhile,the association between SNPs and the risk of ASD was checked in our previous Genome-wide association study（GWAS） database of congenital septation defects. Results：Rs1044598mutant genotype AA decreased the risk of ASD by 36% compared with wild genotype TT. The luciferase reporter assay further confirmed that the significant differences were detected in cotransfected plasmid（rs1044598） with mi R-4252 in HEK293 T,H9C2 and cardiac cells from newborn SD rats cell lines（all P 〈0.05）. Conclusion：Rs1044598 may be a new independent susceptibility locus of ASD. Mi R-4252 down-regulates COL6A2 expression by binding to 3′-UTR of COL6A2,and rs1044598 could influence the procedure and decrease the risk of ASD.