Association And Transcriptional Activity Of Rs13137343 Mutation In SLC2A9 Gene With Uric Acid And Hyperuricemia

Objective:The aim of this study was to explore the association between rs13137343polymorphism in the promoter region of SLC2A9 gene and serum uric acid and hyperuricemia in Xinjiang population,and to detect rs13137343 locus using site-directed mutagenesis and dual luciferase reporter gene system.The influence of gene transcriptional activity provides new ideas and new targets for the research and drug development of hyperuricemia in this region.Methods:From March 2017 to May 2018,1287 patients with hyperuricemia between the ages of 20 and 69 years and 1739 patients with healthy controls were recruited according to inclusion and exclusion criteria.The subjects were examined by standard methods.Blood biochemical indicators were tested,and the basic clinical indicators of the subjects and the prevalence of hyperuricemia were analyzed;the physical examination and blood biochemical index were tested by standard method,and the basic clinical indexes and the prevalence of hyperuricemia were analyzed.The rs13137343 locus of SLC2A9 gene was genotyped by multiplex high temperature ligase assay（iMLDR）.The genotypes and allelic frequencies were analyzed by case-control study and stratified analysis.The effect of rs13137343 site mutation on the transcriptional activity of SLC2A9 gene was detected by dual-luciferase reporter system.Results:The study found that the SBP,DBP,SUA,FBG,TG and BUN in the hyperuricemia group were higher than those in the control group,and the TC and HDL-C were lower than the control group（P<0.05）.The incidence of hyperuricemia in men before the age of 50 was higher than that in women,and that women were higher than men after age 50（P<0.05）.There was no significant difference in genotype frequency of SLC2A9 rs13137343 locus between hyperuricemia group and control group（P=0.052）,but the allele frequency distribution was statistically significant（P<0.05）,the frequency of the A allele in the hyperuricemia group was 51.9%,the frequency in the control group was 49.2%,and the frequency of the C allele in the hyperuricemia group was 48.1%in the control group.Individuals carrying A allele had a higher risk of developing hyperuricemia（OR=1.111,95%CI=1.004¹.231）than those with C allele.There was no significant difference in genotype and allele frequency of SLC2A9 gene rs13137343 between male hyperuricemia group and control group(c²_Genotype=0.855,P=0.652;c²_Allele=0.073,P=0.787),but there was a statistically significant difference between the female hyperuricemia group and the control group(c²_Genotype=9.560,P=0.008;c²_Allele=8.778,P=0.003).The frequency of A allele in the hyperuricemia group（54.7%）was higher than that in the control group（48.4%）.Individuals with A allele are 1.287 times more likely to develop hyperuricemia than those who do not（OR=1.287,95%CI=1.089¹.522）.The genotype and allele frequency of this locus in the hyperuricemia group of different sexes showed that the frequency of female A allele was 54.7%higher than that of male（50.1%）,and the frequency of C allele was 45.3%lower than that of male（49.9%）,the difference was statistically significant(c²_Allele=5.163,P=0.023),but there was no significant difference in genotype and allele frequency distribution between male and female in the control group（P>0.05）.Women with AA genotype had 1.619 times higher risk of hyperuricemia than those with CC genotype,after adjusting for age,it was found that the AA genotype of this locus was still associated with the risk of hyperuricemia in women.（95%CI=1.154².237）,suggesting that women carrying the AA genotype will increase the risk of developing hyperuricemia;There was a significant difference in the frequency of AA vs CC+AC genotype between the hyperuricemia group and the control group（P<0.05）.The frequency of AA genotype in the case group was higher than that of the control group,and the women with AA genotype had higher risk of hyperuricemia（female OR=1.495,95%CI=1.148¹.948）;Women with AA genotype had a higher risk of hyperuricemia（female OR=1.495,95%CI=1.148¹.948）.After adjusting for age,the AA genotype of this locus was still higher than that of women.The risk of uric acidemia was associated with OR=1.607（95%CI=1.154-2.237）,suggesting that women with AA genotypes may increase the risk of developing hyperuricemia.There was no significant difference in genotype and allele frequency distribution of rs13137343 locus of SLC2A9gene in male with different uric acid level,but there was significant difference in female（P<0.05）,and the frequency of A allele increased with the increase of uric acid level,and the frequency of C allele decreased with the increase of uric acid level.The site-directed mutagenesis technique and the dual luciferase reporter gene system were used to detect whether the rs13137343 site mutation affects the SLC2A9 gene promoter activity.The results showed that the luciferase activity was down regulated by 42.51%,suggesting that the site has an important regulatory effect on the transcriptional activity of SLC2A9 gene.Conclusion:（1）The rs13137343 locus of SLC2A9 gene was associated with the occurrence of hyperuricemia in the population of Xinjiang,and was more closely related to the occurrence of hyperuricemia in women.The A allele has a positive effect on the increase of uric acid level compared with the C allele,considering the mutation of C→A leads to disorder of uric acid metabolism and the risk of hyperuricemia is increased.（2）The site-directed mutagenesis technique and the dual luciferase reporter gene system revealed that the rs13137343 site mutation played an important role in regulating the transcriptional activity of SLC2A9 gene.