Division of Personalized Nutrition and Medicine, National Center for Toxicological Research, Food and Drug Administration, Jefferson, AR 72079, USA.
J Biochem Mol Toxicol. 2012 Oct;26(10):422-8. doi: 10.1002/jbt.21437.
Previously, we reported five common single nucleotide polymorphisms (SNPs), -624G>C, -396G>A, -358A>C, -341C>G, and -294T>C, and six common haplotypes (CGACT, GAACT, GGAGC, GGACC, CAACT, and GAACC) in the 5'-flanking region of the SULT1A1 gene that were associated with altered enzymatic activity. In the present study, we performed in vitro assays to determine the functional impact of these genetic variations on the promoter activity. Dual luciferase reporter assays revealed that these SNPs are located in a negative regulatory fragment of the SULT1A1 gene. Further experiments demonstrated that these SNPs and haplotypes affected promoter activities of SULT1A1. Electrophoretic mobility shift assays showed distinctive binding patterns for the SNPs -396G>A and -294T>C, due to differential binding affinities of the G/A alleles and the T/C alleles to nuclear proteins extracted from the liver carcinoma cell lines, HepG2 and Huh7.
此前，我们报道了 SULT1A1 基因 5' 侧翼区的五个常见单核苷酸多态性（SNPs），-624G>C、-396G>A、-358A>C、-341C>G 和 -294T>C，以及六个常见单倍型（CGACT、GAACT、GGAGC、GGACC、CAACT 和 GAACC），这些与酶活性改变有关。在本研究中，我们进行了体外实验来确定这些遗传变异对启动子活性的功能影响。双荧光素酶报告基因检测显示，这些 SNPs 位于 SULT1A1 基因的负调控片段中。进一步的实验表明，这些 SNPs 和单倍型影响了 SULT1A1 的启动子活性。电泳迁移率变动分析显示，由于 G/A 等位基因和 T/C 等位基因与肝癌细胞系 HepG2 和 Huh7 中提取的核蛋白的结合亲和力不同，SNP-396G>A 和 -294T>C 存在不同的结合模式。
