PDZK1基因启动子区rs12129861位点基因多态性与新疆汉族男性人群痛风发病的关系

Relationship between gene polymorphism of the rs12129861 locus in the promoter region of PDZK1 gene and the incidence of gout in Xinjiang Han male population

目的了解PDZ蛋白激酶1(PDZK1)基因启动子区rs12129861(A/G)位点的基因型和基因频率与新疆地区汉族男性人群痛风发病的关系。方法收集汉族男性痛风患者(痛风组)和同时期体检健康者(对照组)的血样,采用多重荧光PCR技术检测PDZK1基因启动子区rs12129861(A/G)位点的基因型。比较两组rs12129861(A/G)位点的基因型分布及基因频率以及不同rs12129861(A/G)位点的基因型人群血清尿酸浓度。选择KpnI、XhoI双酶切位点,构建含rs12129861(A/G)位点的pGL3-promotor荧光素酶表达载体,采用双荧光素酶报告基因实验证实启动子区rs12129861(A/G)位点是否影响荧光素酶基因的表达。结果PDZK1基因启动子区rs12129861(A/G)位点基因型分布符合Hardy-Weinberg平衡性检验(P>0.05),该位点稳定遗传具有群体代表性。痛风组和对照组PDZK1基因启动子区rs12129861(A/G)位点AA、GA、GG基因型分布频率差异有统计学意义(χ^(2)=14.633,P<0.01),等位基因A和G分布频率的差异有统计学意义(χ^(2)=14.236,P<0.01),其中痛风组等位基因G的分布频率高于对照组,OR值为1.661。rs12129861(A/G)位点基因型为A/A与G/G人群间、G/A与G/G人群间血清尿酸浓度比较,P<0.05,其中基因型为G/G的人群血尿酸浓度大于基因型A/A、G/A人群。双荧光素酶实验结果显示,rs12129861(A/G)具有增强子功能,rs12129861(G)重组质粒的荧光素酶报告基因表达水平低于rs12129861(A)重组质粒(P<0.05)。结论PDZK1基因启动子区rs12129861位点的基因型和基因频率与新疆汉族男性痛风有关,等位基因G可能是痛风发病的危险因素。

Objective To investigate the association between the genotype and gene frequency of the rs12129861(A/G)locus in the promoter region of the PDZ protein kinase 1(PDZK1)gene and the incidence of gout in Xinjiang Han male population.Methods Blood samples of Han male gout patients(gout group)and healthy subjects(control group)at the same period were collected,and the genotypes of rs12129861(A/G)locus in PDZK1 gene promoter region were detected by multi-fluorescence PCR.The genotype distribution and gene frequency of rs12129861(A/G)locus and serum uric acid concentration of population with different genotypes at rs12129861(A/G)locus were compared between the two groups.KpnI and XhoI double digestion sites were selected to construct the pGL3-promotor luciferase expression vector containing rs12129861(A/G)site.Double luciferase reporter gene assay was used to confirm whether rs12129861(A/G)site in the promoter region affected luciferase gene expression.Results The genotype distribution of rs12129861(A/G)locus in the promoter region of PDZK1 gene was consistent with Hardy-Weinberg balance test(P>0.05),and the stable inheritance of this locus was population representative.There were statistically significant differences in the distribution frequencies of AA,GA and GG genotypes at rs12129861(A/G)locus in PDZK1 gene promoter region between the gout group and control group(χ^(2)=14.633,all P<0.01),the difference of allele A and G distribution frequency was statistically significant(χ^(2)=14.236,P<0.01),the distribution frequency of allele G in the gout group was higher than that in the control group,and the OR value was 1.661.Significant difference was found in the serum uric acid concentration between the population with genotype A/A and genotype G/G,and between the population with genotype G/A and genotype G/G at rs12129861(A/G)locus(both P<0.05).The serum uric acid concentration of G/G genotype was higher than those of A/A genotype and G/A genotype.The results of double luciferase assay showed that rs12129861(A/G)had enhancer function,and the luciferase reporter expression level of rs12129861(G)was lower than that of rs12129861(A)(P<0.05).Conclusion The genotype and gene frequency of rs12129861 in the promoter region of PDZK1 gene are associated with gout in Xinjiang Han male population,and allele G may be a risk factor for gout.