2型糖尿病病人中SIRT2基因启动子遗传变异和功能研究

发布时间：2018-04-17 00:27

本文选题：2型糖尿病 + SIRT2　；参考：《济宁医学院》2017年硕士论文

【摘要】：目的:通过对2型糖尿病(Type 2 diabetes mellitus,T2DM)病人和对照组人群的SIRT2基因启动子序列进行遗传学和功能分析,找出SIRT2基因启动子序列变异与T2DM发生的关系。方法:本实验运用大样本病例对照研究,共纳入T2DM病人365例,对照人群358例。采用聚合酶链反应的方法,对SIRT2基因启动子序列进行扩增,后DNA测序,与野生型对比分析,以发现两组人群中SIRT2基因启动子的DNA序列变异情况。通过双荧光素酶报告分析对DNA序列变异进行功能分析。将变异的SIRT2基因启动子进行扩增,再插入pGL3-Basic载体,转染胰腺细胞β-1和人肾胚-293细胞,测量双荧光素酶活性,通过两组之间对比分析DNA序列变异对SIRT2基因启动子转录活性的影响。结果:本研究两组人群中共发现18个DNA序列变异(DNA sequence variants,DSVs),包括5个单核苷酸多态性(single-nucleotide polymorphisms,SNPs)。我们在T2DM病人组中,发现3个为新颖杂合DSVs,g.38900912GT,g.38900359CT,g.38900237GA(在对照组中没有发现)。在体外培养的细胞中,上述DSVs显著增加SIRT2基因启动子的转录活性。只在对照组中发现的7个新杂合子DSVs,以及在病人组中和对照组中均出现的一个杂合缺失和5个单核苷酸多态性,它们均未显著改变SIRT2基因启动子活性。结论:我们的发现提示,SIRT2基因启动子的DNA序列变异可能通过影响了启动子转录活性,影响SIRT2基因表达水平,作为危险因素参与T2DM的发生。
[Abstract]:Objective: to study the genetic and functional analysis of SIRT2 gene promoter sequence in type 2 diabetes mellitus patients with type 2 diabetes mellitusus T2DM and control group, and to find out the relationship between SIRT2 gene promoter mutation and T2DM.Methods: a large sample case-control study was conducted in 365 T2DM patients and 358 controls.Polymerase chain reaction (PCR) was used to amplify the promoter sequence of SIRT2 gene, then DNA sequence was sequenced, and compared with wild type, the variation of DNA sequence of promoter of SIRT2 gene in two groups was found.The variation of DNA sequence was analyzed by double luciferase report analysis.The mutated SIRT2 gene promoter was amplified and inserted into pGL3-Basic vector, then transfected into pancreatic 尾 -1 and human renal embryo 293 cells. The activity of double luciferase was measured. The effect of DNA sequence mutation on the transcriptional activity of SIRT2 gene promoter was compared between the two groups.Results: a total of 18 DNA variants were found in the two groups, including 5 single-nucleotide polymorphisms (SNPs) and 5 single-nucleotide polymorphisms (SNPs).In the T2DM patient group, we found three novel heterozygous DSVsN. 38900912 GTG. 38900359CTL. 38900237GA (not found in the control group).In vitro cultured cells, DSVs significantly increased the transcriptional activity of SIRT2 gene promoter.Only 7 new heterozygotes found in the control group, one heterozygosity deletion and five single nucleotide polymorphisms in the patient group and the control group did not significantly change the promoter activity of the SIRT2 gene.Conclusion: our findings suggest that the DNA sequence variation of SIRT2 gene promoter may affect the transcriptional activity of the promoter and the level of SIRT2 gene expression, which may be a risk factor in the pathogenesis of T2DM.
【学位授予单位】：济宁医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R587.1

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