MiR-126和miR-21基因多态性与缺血性脑卒中的遗传易感性研究

发布时间：2018-05-10 15:19

本文选题：miR-126 + 基因　；参考：《右江民族医学院》2017年硕士论文

【摘要】：目的:检测广西地区健康人群miR-126基因rs4636297G/A多态性位点的分布频率,并与其他不同地区人群rs4636297G/A位点的分布频率进行比较,揭示广西地区健康人群rs4636297G/A位点的分布特点。方法:采用单碱基延伸的聚合酶链式反应(Snapshot PCR)基因分型技术和DNA测序的方法来检测广西地区456例健康人miR-126基因rs4636297G/A的多态性,统计rs4636297G/A位点的基因型和等位基因的频率分布;通过Pub Med(SNP)数据库获取人类基因组计划公布的中国北京人群、欧洲人群、日本人群及非洲人群的rs4636297G/A位点分型数据;将广西地区人群rs4636297G/A多态性位点分型数据与其他四个地区人群的分型数据进行比较,来分析这五个地区人群的多态性位点频率分布的异同。结果:根据基因分型技术和DNA测序的方法检测的结果可知,广西地区健康人群miR-126基因rs4636297G/A位点存在多态性,并具有三种基因型,且多态性性分布均符合Hardy-Weinberg遗传平衡定律(P0.05),具有明显的群体代表性。将广西地区miR-126基因rs4636297G/A位点的分型数据与其他四个地区的分型数据比较发现,miR-126基因rs4636297G/A位点在广西人群、北京和日本人群间多态性的分布无显著性差异(P均0.05),而miR-126基因rs4636297G/A位点在广西人群和其他两个人群间的频率分布存在显著性的差异(P均0.05)。结论:在广西地区健康人群中存在miR-126基因rs4636297G/A位点多态性,且多态性分布均符合Hardy-Weinberg遗传平衡定律(P0.05),说明具有明显的群体代表性。MiR-126基因rs4636297G/A位点多态性的分布在广西地区和其他地区人群不完全一致,这可能是不同地区人群对疾病的易感性表现出差异的原因之一。目的:探讨miR-126基因rs4636297G/A多态性与缺血性脑卒中发生的遗传易感性,检测缺血性脑卒中患者与正常人群中miR-126的表达水平,并分析miR-126基因rs4636297G/A多态性与miR-126的表达是否存在相关性。方法:采用单碱基延伸的聚合酶链式反应(Snapshot PCR)基因分型技术和DNA测序的方法来检测广西地区456例健康人和592例缺血性脑卒中患者miR-126基因rs4636297G/A多态性位点的多态性,计算rs4636297G/A多态性位点在健康对照组及病例组中的基因型和等位基因的频率分布,利用统计软件分析miR-126基因rs4636297G/A多态性与缺血性脑卒中发生的遗传易感性。利用SYBR Green实时荧光定量PCR的方法检测病例对照组中miR-126的表达水平,并分析miR-126基因rs4636297G/A多态性与miR-126表达的相关性。结果:MiR-126基因rs4636297G/A位点在病例和对照组中都存在多态性,分别具有三种基因型,且rs4636297G/A多态性位点在病例对照组中的频率分布均符合Hardy-Weinberg遗传平衡定律(P均0.05),具有明显的群体代表性。在本次研究中我们发现miR-126基因rs4636297G/A位点的频率分布在缺血性脑卒中病人及健康人群间存在显著性差异(P0.05)。与健康人群相比较,缺血性脑卒中患者GA(33.8%vs.24.8%)和AA(5.9%vs.2.5%)基因型的频率显著降低,GA和AA基因型与缺血性脑卒中的发生负相关(GA vs.GG:OR=0.61,95%CI=0.47-0.80,P0.001;AA vs.GG:OR=0.36,95%CI=0.19-0.68,P=0.001);进行遗传模型分析后发现miR-126基因rs4636297G/A位点的显性模型和隐性模型在病例对照组间均存在显著差异(显性模型,AA+GA vs.GG:OR=0.57,95%CI=0.44-0.74,P0.001;隐性模型,AA vs.GA+AA:OR=0.41,95%CI=0.22-0.79,P=0.006)。通过纳入缺血性脑卒中的常见风险因素,年龄、性别、吸烟、高血压、糖尿病、甘油三酯、胆固醇、低密度脂蛋白及高密度脂蛋白进行logistic回归校正计算后发现,miR-126基因rs4636297G/A多态性位点的基因型及遗传模型的分布在病例对照间仍然存在显著性的差异(GA vs.GG:AOR=0.64,95%CI=0.44-0.95,P=0.025;AA vs.GG:AOR=0.32,95%CI=0.14-0.74,P=0.007;显性模型,AA+GA vs.GG:AOR=0.58,95%CI=0.40-0.84,P=0.004;隐性模型,AA vs.GA+AA:AOR=0.37,95%CI=0.16-0.82,P=0.015)。与正常组对比,缺血性脑卒中病人中miR-126的表达显著降低(缺血性脑卒中组vs.对照组=6.57±1.50 vs.9.86±1.92,P0.001),携带GA/AA基因型的患者其miR-126表达水平显著高于携带GG患者的miR-126表达水平(GG vs.GA/AA=6.08±1.47 vs.7.53±2.00,P0.001);正常对照组中rs4636297G/A多态性与miR-126的表达不存在相关性(GG vs.GA/AA=9.77±1.67 vs.10.6±1.19,P=0.485)。结论:MiR-126基因rs4636297G/A位点GA/AA基因型频率在缺血性脑卒中患者中显著减少,与缺血性脑卒中的发生存在负相关性,miR-126基因rs4636297G/A位点可能是缺血性脑卒中发生的一个保护性的因素。此外,缺血性脑卒中患者中异常减少miR-126可能是其潜在的生物学标记或者治疗靶标。目的:检测广西地区健康人群miR-21基因rs1292037T/C多态性位点的分布频率,并与不同地区人群进行比较,揭示广西地区rs1292037T/C多态性位点的分布特点,为后续的关联性研究奠定遗传学基础。方法:采用单碱基延伸的聚合酶链式反应(Snapshot PCR)基因分型技术和DNA测序的方法来检测广西地区456例健康人群rs1292037T/C多态性位点的多态性,统计这个位点的基因型和等位基因的频率分布;通过Pub Med(SNP)数据库来获取人类基因组计划公布的中国北京人群、欧洲人群、日本人群及非洲人群的SNPs分型数据;将广西地区人群的rs1292037T/C多态性位点分型数据与其他四个地区人群的分型数据进行比较,来分析这五个地区人群的多态性位点频率分布的异同。结果:根据基因分型技术和DNA测序的方法检测的结果可知,广西地区人群miR-21基因rs1292037T/C位点存在多态性,并且具有三种基因型,rs1292037T/C的多态性分布符合Hardy-Weinberg遗传平衡定律(P0.05),具有明显的群体代表性。将广西地区miR-21基因rs1292037T/C位点的分型数据与其他四个地区的分型数据比较发现,miR-21rs1292037T/C位点在广西人群和北京人群间的频率分布无显著性差异(P0.05),而rs1292037T/C位点在广西人群和其他地区三个人群间的频率分布存在显著性的差异(P均0.05)。结论:在广西地区人群中存在miR-21基因rs1292037T/C多态性,rs1292037T/C多态性分布符合Hardy-Weinberg遗传平衡定(P0.05),具有明显的群体代表性。miR-21基因rs1292037T/C多态性的分布在广西地区和其他地区人群不一致,这可能是不同地区人群对疾病易感性表现出差异的原因之一。目的:探讨miR-21基因rs1292037T/C多态性与缺血性脑卒中发生的遗传易感性,检测缺血性脑卒中患者与正常人群中miR-21的表达水平,并分析miR-21基因rs1292037T/C多态性与miR-21的表达是否存在相关性。方法:采用单碱基延伸的聚合酶链式反应(Snapshot PCR)基因分型技术和DNA测序的方法来检测广西地区456健康对照人群和592例缺血性脑卒中患者miR-21基因rs1292037T/C位点的多态性,计算rs1292037T/C多态性位点在健康对照组及病例组中的基因型和等位基因的频率分布,通过统计软件来评估miR-21基因rs1292037T/C多态性与缺血性脑卒中发生的遗传易感性。利用SYBR Green实时荧光定量PCR的方法检测病例对照组中miR-21的表达水平,并分析miR-21基因rs1292037T/C多态性与miR-21表达的相关性。结果:MiR-21基因rs1292037T/C位点在病例和对照组中都存在多态性,rs1292037T/C位点具有三种基因型,rs1292037T/C多态性位点在病例对照组中的频率分布均符合Hardy-Weinberg遗传平衡定律(P均0.05),具有明显的群体代表性。在本次研究中我们发现rs1292037T/C多态性位点的频率分布在缺血性脑卒中病人及健康人群间不存在显著性差异(P0.05)。进行遗传模型分析后发现miR-21基因rs1292037T/C位点显性模型和隐性模型在病例对照组间也不存在显著差异(显性模型,CC+CT vs.TT:OR=1.22,95%CI=0.94-1.59,P=0.137;隐性模型,CC vs.CT+TT:OR=1.24,95%CI=0.90-1.71,P=0.191)。通过纳入缺血性脑卒中的常见风险因素如年龄、性别、吸烟、高血压、糖尿病、甘油三酯、胆固醇、低密度脂蛋白及高密度脂蛋白进行logistic回归校正计算后发现,rs1292037T/C多态性位点的基因型及遗传模型的分布在病例对照间的分布仍然不存在显著性的差异(CT vs.TT:AOR=1.21,95%CI=0.81-1.80,P=0.349;CC vs.TT:AOR=1.47,95%CI=0.86-2.50,P=0.159;显性模型,CC+CT vs.TT:OR=1.28,95%CI=0.88-1.86,P=0.204;隐性模型,CC vs.CT+TT:OR=1.37,95%CI=0.85-2.21,P=0.192),提示rs1292037T/C多态性可能与缺血性脑卒中的发生无相关性。在缺血性脑卒中病人中miR-21的表达显著升高(缺血性脑卒中组vs.对照组=34.65±7.26 vs.26.89±6.81,P0.001),但miR-21的表达水平与rs1292037T/C多态性无关。结论:MiR-21基因rs1292037T/C多态性与缺血性脑卒中的发生不存在相关性,缺血性脑卒中患者中异常增高的miR-21可能是其潜在的生物学标记和治疗靶标。
[Abstract]:Objective: to detect the distribution frequency of miR-126 gene rs4636297G/A polymorphic loci in healthy people in Guangxi, and to compare the distribution frequency of rs4636297G/A loci with other people in other regions, to reveal the distribution of rs4636297G/A loci in healthy people in Guangxi. Method: single base extension polymerase chain reaction (Snapshot PCR). Genotyping and DNA sequencing methods were used to detect the polymorphism of miR-126 gene rs4636297G/A in 456 healthy people in Guangxi, and the frequency distribution of genotypes and alleles of the rs4636297G/A loci were statistically analyzed. The human genome project, the European population, the Japanese population and the Africans, were obtained by the Pub Med (SNP) database. The rs4636297G/A polymorphism data of the group were compared with the typing data of the rs4636297G/A polymorphic loci in the Guangxi population and the other four regions, to analyze the similarities and differences in the frequency distribution of polymorphic loci in the five regions. The miR-126 gene rs4636297G/A loci of the healthy people in the western region have polymorphism and have three genotypes, and the polymorphic distribution conforms to the Hardy-Weinberg's law of genetic balance (P0.05). It has a distinct population representation. The classification data of the rs4636297G/A loci of the miR-126 gene in Guangxi area are compared with the other four regions. It was found that there was no significant difference in the distribution of polymorphism between the miR-126 gene rs4636297G/A loci in Guangxi population, Beijing and the Japanese group (P 0.05), while the frequency distribution of miR-126 gene rs4636297G/A locus in Guangxi population and other two groups was significantly different (P 0.05). Conclusion: there is miR-126 in the healthy population of Guangxi. The polymorphism of the gene rs4636297G/A locus and the polymorphism distribution conform to the Hardy-Weinberg's law of genetic balance (P0.05), indicating that the distribution of the rs4636297G/A locus polymorphism of the group representative.MiR-126 gene is not exactly the same in the Guangxi region and other populations. This can be the expression of the susceptibility to disease in different regions. Objective: To investigate the genetic susceptibility of miR-126 gene rs4636297G/A polymorphism to ischemic stroke, to detect the expression level of miR-126 in ischemic stroke patients and normal people, and to analyze the correlation between the miR-126 gene rs4636297G/A polymorphism and the expression of miR-126. Method: single base extension is used. Polymerase chain reaction (Snapshot PCR) genotyping and DNA sequencing were used to detect the polymorphism of miR-126 gene polymorphism loci in 456 healthy and 592 ischemic stroke patients in Guangxi, and to calculate the genotype and allele frequency of rs4636297G/A polymorphic loci in the healthy control group and the case group. Rate distribution, the miR-126 gene rs4636297G/A polymorphism and the genetic susceptibility to ischemic stroke were analyzed by statistical software. The expression level of miR-126 in the case control group was detected by SYBR Green real-time fluorescent quantitative PCR, and the correlation between the miR-126 gene rs4636297G/A polymorphism and miR-126 expression was analyzed. The result: MiR-126 gene. Rs4636297G/A loci have polymorphism in both cases and control groups, with three genotypes, respectively, and the frequency distribution of rs4636297G/A polymorphic loci in case control group conforms to Hardy-Weinberg's law of genetic balance (P 0.05), which has obvious population representation. In this study, we found the rs4636297G/A locus of the miR-126 gene. There was a significant difference in frequency distribution between ischemic stroke patients and healthy people (P0.05). Compared with healthy people, the frequency of GA (33.8%vs.24.8%) and AA (5.9%vs.2.5%) genotypes in ischemic stroke patients decreased significantly, and GA and AA genotypes were negatively correlated with ischemic stroke (GA vs.GG:OR=0.61,95%CI=0.47-0.80, P0.001). AA vs.GG:OR=0.36,95%CI=0.19-0.68, P=0.001); after the analysis of the genetic model, the dominant and recessive models of the miR-126 gene rs4636297G/A locus were significantly different between the case control groups (dominant model, AA+GA vs.GG:OR=0.57,95%CI=0.44-0.74, P0.001; implicit model, AA vs.GA+AA:OR=0.41,95%CI=0.22-0.79, P=0.006). Logistic regression analysis of the common risk factors for ischemic stroke, age, sex, smoking, hypertension, diabetes, triglycerides, cholesterol, low density lipoprotein and HDL, found that the genotype and genetic model of the miR-126 gene rs4636297G/A polymorphism still remained in case control. In significant differences (GA vs.GG:AOR=0.64,95%CI=0.44-0.95, P=0.025; AA vs.GG:AOR=0.32,95%CI=0.14-0.74, P=0.007; dominant model, AA+GA vs.GG:AOR=0.58,95%CI=0.40-0.84, P=0.004; recessive model, AA vs.GA+AA:AOR=0.37,95%CI=0.16-0.82, P=0.015). In the ischemic stroke group, the vs. control group was =6.57 + 1.50 vs.9.86 + 1.92, P0.001). The level of miR-126 expression in the patients carrying GA/AA genotype was significantly higher than that of the GG patients (GG vs.GA/AA=6.08 + 1.47 vs.7.53 + 2, P0.001), and there was no correlation between the normal control group and the expression of the normal control group. A=9.77 + 1.67 vs.10.6 + 1.19, P=0.485). Conclusion: the frequency of GA/AA genotype of rs4636297G/A locus of MiR-126 gene is significantly reduced in ischemic stroke patients and has a negative correlation with the occurrence of ischemic stroke. The rs4636297G/A site of miR-126 gene may be a protective factor for ischemic stroke. In addition, ischemic brain Abnormal reduction of miR-126 in stroke patients may be a potential biological marker or therapeutic target. Objective: to detect the distribution frequency of rs1292037T/C polymorphic loci of the miR-21 gene in healthy people in Guangxi and to compare the distribution of the rs1292037T/C polymorphic loci in the Guangxi region. The genetic basis of sex study was established. Methods: single base extended polymerase chain reaction (Snapshot PCR) genotyping and DNA sequencing were used to detect polymorphism of rs1292037T/C polymorphic loci in 456 healthy people in Guangxi region, and the frequency distribution of genotypes and alleles of this site was statistically analyzed; the number of Pub Med (SNP) numbers was measured. The SNPs classification data of the Chinese Beijing population, the European population, the Japanese population and the African population were obtained from the human genome project, and the rs1292037T/C polymorphic loci data of the Guangxi population were compared with the subtypes of the other four regions to analyze the frequency of polymorphic loci in the five regions. Results: according to the results of genotyping and DNA sequencing, the rs1292037T/C loci of miR-21 gene in Guangxi population are polymorphic and have three genotypes. The polymorphism distribution of rs1292037T/C conforms to the Hardy-Weinberg's law of genetic balance (P0.05). It has a distinct population representation. It will be the place of Guangxi. Compared with the typing data of the other four regions, the rs1292037T/C loci of the region miR-21 gene showed that there was no significant difference in the frequency distribution of the miR-21rs1292037T/C loci between the Guangxi population and the Beijing population (P0.05), while the frequency distribution of the rs1292037T/C locus between the Guangxi population and the other three populations had a significant difference. Difference (P 0.05). Conclusion: there is a rs1292037T/C polymorphism of miR-21 gene in the population of Guangxi, and the distribution of rs1292037T/C polymorphism conforms to Hardy-Weinberg genetic balance (P0.05). The distribution of rs1292037T/C polymorphism of the group representative.MiR-21 gene is not consistent with those in other regions, which may be different regions. One of the reasons for the difference in susceptibility to disease. Objective: To investigate the genetic susceptibility of miR-21 gene rs1292037T/C polymorphism to ischemic stroke, to detect the expression level of miR-21 in ischemic stroke patients and normal population, and to analyze the correlation between the rs1292037T/ C polymorphism of miR-21 gene and the expression of miR-21. Methods: single base extended polymerase chain reaction (Snapshot PCR) genotyping and DNA sequencing were used to detect the polymorphism of miR-21 gene rs1292037T/C loci in 456 healthy controls and 592 patients with ischemic stroke in Guangxi area, and to calculate the genes of rs1292037T/C polymorphic loci in healthy control and case groups. The frequency distribution of the genotype and allele was used to evaluate the genetic susceptibility of the miR-21 gene rs1292037T/C polymorphism to the ischemic stroke. The expression of miR-21 in the case control group was detected by SYBR Green real-time fluorescent quantitative PCR, and the correlation between the miR-21 based rs1292037T/C polymorphism and the miR-21 expression was analyzed. Results: MiR-21 gene rs1292037T/C loci were polymorphic in both cases and control groups, and there were three genotypes in rs1292037T/C loci. The frequency distribution of rs1292037T/C polymorphic loci in case control group all conformed to Hardy-Weinberg's law of genetic balance (P 0.05). The frequency distribution of the present rs1292037T/C polymorphic loci was not significant difference between the ischemic stroke patients and the healthy people (P0.05). After the genetic model analysis, it was found that there was no significant difference between the dominant and recessive model of the miR-21 gene rs1292037T/C locus in the case control group (the dominant model, CC+CT vs.TT:OR=1.22,95%CI=0.). 94-1.59, P=0.137; recessive model, CC vs.CT+TT:OR=1.24,95%CI=0.90-1.71, P=0.191). The rs1292037T/C polymorphism was found by logistic regression analysis of common risk factors included in ischemic stroke such as age, sex, smoking, hypertension, diabetes, triglycerides, cholesterol, low density lipoprotein and high density lipoprotein. The distribution of point genotypes and genetic models still had no significant difference between the distribution of case control (CT vs.TT:AOR=1.21,95%CI=0.81-1.80, P=0.349; CC vs.TT:AOR=1.47,95%CI=0.86-2.50, P=0.159; dominant model, CC+CT vs.TT:OR=1.28,95%CI=0.88-1.86, P=0.204; recessive model, CC vs.CT+TT:OR=1.37,95%CI=0.85-2.21, P=0.192), It is suggested that rs1292037T/C polymorphism may not be associated with the occurrence of ischemic stroke. The expression of miR-21 in ischemic stroke patients is significantly elevated (=34.65 + 7.26 vs.26.89 + 6.81, P0.001) in ischemic stroke group (vs. control group), but the expression level of miR-21 is not related to rs1292037T/C polymorphisms. Conclusion: MiR-21 gene rs1292037T/C polymorphism There is no correlation between the occurrence of ischemic stroke and the occurrence of ischemic stroke. The abnormally high miR-21 in ischemic stroke may be its potential biological marker and therapeutic target.

【学位授予单位】：右江民族医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R743.3

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