人胰岛素样生长因子-I基因P1启动子区域单核苷酸多态性-705T>C和-603T>A对启动子活性的影响(英文)

目的探讨人胰岛素样生长因子-I(IGF-I)基因P1启动子区域单核苷酸多态性(SNP)-705T>C和-603T>A对启动子活性的影响。方法招募152名健康志愿者参与本研究,取静脉血样品,使用基因组DNA提取试剂盒从血样中提取全基因组DNA。通过限制片段长度多态性分析(RFLP),对每名志愿者的SNP-705T>C和-603T>A进行基因分型,统计各基因型的频率。使用PHASE v2.1软件程序分析P1启动子单体型的类型和频率。通过荧光素酶报告基因测定不同P1启动子单体型的活力差异。结果成功提取了每名志愿者的全基因组DNA并测定了SNP-705T>C和-603T>A的基因型。基因型-705T/T、705T/C、-705C/C的频率分别与-603T/T、-603T/A、-603A/A相同,分别为43.4%、45.4%、11.2%。等位基因-705T、-705C的频率分别与-603T、-603A相同,分别为66.1%、33.9%。SNP-705T>C和-603T>A之间存在完全的连锁不平衡,其组成的P1启动子单体型只有-705T/-603T和-705C/-603A两种,频率分别为66.1%和33.9%。报告基因分析结果表明,P1启动子-705C/-603A单体型的活力显著高于-705T/-603T单体型。结论人IGF-I基因P1启动子区域SNP-705T/C和-603T/A能够影响P1启动子的活性。

Effect of single-nucleotide polymorphisms-705T/C and-603T/A in P1 promoter region of human insulin-like growth factor-I Gene on promoter activity

Objective The present study aims to investigate the effect of single-nucleotide polymorphisms(SNP)-705T>C and-603T>A in the P1 promoter region of human insulin-like growth factor I(IGF-I) gene on promoter activity.Methods A total of 152 healthy volunteers were recruited for the present study.The peripheral blood samples were obtained through venipuncture.The total genomic DNA of each blood sample was extracted using the genomic DNA extraction kit.Genotyping of SNP-705T>C and-603T>A in each volunteer was performed based on restriction fragment length polymorphism(RFLP) analysis,and the frequencies of each genotype were statistically analyzed.P1 promoter haplotypes and their frequencies were analyzed by PHASE v2.1 software program.Luciferase reporter gene assays were adopted to determine the difference among activity of different promoter haplotypes.Results The total genomic DNA of each volunteer was successfully extracted.Genotyping of SNP-705T>C and-603T>A was also carried out.The genotype frequencies of-705T/T,-705T/C,and-705C/C were the same as those of-603T/T,-603T/A,and-603A/A,which were 43.4%,45.4%,and 11.2%,respectively.The frequencies of allelic gene-705T and-705C were the same as those of-603T and-603A,which were 66.1% and 33.9%,respectively.A complete linkage disequilibrium between SNP-705T>C and-603T>A was observed.Moreover,these two SNPs only comprised two types of promoter haplotypes,including-705T/-603T(66.1%) and-705C/-603A(33.9%).The results of reporter gene assays showed that the activity of-705C/-603A P1 promoter haplotype was significantly higher than that of-705T/-603T haplotype.Conclusion SNP-705T>C and-603T>A in the P1 promoter field of human IGF-I gene can affect the activity of the P1 promoter.