DNMT1、KIT和KITLG基因的单核苷酸多态性与生精障碍关系的初步研究
发布时间：2019-06-19 17:47
【摘要】：男性不育是一种由遗传，环境，饮食等诸多因素引起的多基因遗传病。近年来随着不孕不育症发病率不断升高，男性不育也逐渐成为全球最受关注的疾病之一。在人群中，大约有10%至15%育龄夫妇不能正常生育，其中大约有一半是男性不育所致。调查研究表明，男性人群中大约有4%至5%的个体患有遗传因素所导致的男性不育。生精障碍是男性不育最为常见的病因之一，遗传因素在生精障碍的发生过程中起着重要作用。近年来，在生精障碍遗传病因的研究上取得了一定的进展，发现了一些与生精障碍相关的遗传因素，如染色体的畸形，Y染色体的微缺失以及某些基因的突变等。但这些因素只能解释部分的病例，，仍有众多的生精障碍的遗传病因有待于进一步阐明。精子发生是一个复杂的过程，包括精原细胞的增殖分化、减数分裂、精细胞分化成精子等几个阶段。这个过程中有众多基因的参与，这些基因及其表达的异常都有可能导致生精障碍的发生。因此，这些参与生精过程的基因也被认为是生精障碍重要的候选基因，研究这些候选基因与生精障碍的关系，可为男性不育遗传病因、发病机制的最终阐明提供有价值的资料。 DNMT1(DNAmethyltransferase1)基因是近年来发现的一个生精障碍的重要候选基因，其编码产物DNMT1是一种DNA甲基化酶，该酶在DNA的甲基化过程中发挥重要作用。研究表明，精子发生过程中涉及众多基因的表达调控，而DNA甲基化是生精过程中基因表达调控的一种重要方式。某些基因的DNA甲基化是正常生精过程所必须的。在生精过程中，DNMT1基因在细胞增殖、减数分裂、精细胞分化等阶段和成熟的精子中均有表达，主要负责催化和维持染色体复制后的DNA甲基化状态,提示其在正常的生精过程中可能有重要作用。在小鼠中研究显示,DNMT1基因的异常表达,会导致生殖细胞凋亡和生精障碍。在一些生精障碍患者中也发现了DNMT1基因的表达异常。因此,DNMT1基因可能与人类生精障碍有关,其基因变异可能在生精障碍中发挥作用。 近年来,一些信号转导系统与生精障碍的关系也逐渐受到关注。KIT/KITLG信号转导系统被认为是在生殖细胞正常发育调控中不可或缺的一个信号系统。KIT基因编码产物KIT受体是一种酪氨酸激酶受体(tyrosine kinase receptor), KITLG(KIT ligand)基因的产物是KIT受体的特异配体KITLG。该信号系统通过KIT受体与KITLG的特异性结合而激活。在实验动物中研究显示,KIT/KITLG信号转导系统的激活,对睾丸中生殖细胞的增殖、减数分裂、迁移、成熟等十分关键,在生精过程中发挥了重要作用。不同的动物模型研究表明,KIT或KITLG基因的异常,会导致生精障碍和雄性不育。这些研究结果提示,KIT和KITLG基因的变异也可能与人类生精障碍有关。 目前,尚未有DNMT1基因、KIT基因和KITLG基因的变异与人类生精障碍的研究报道。有鉴于此,本研究采用聚合酶链反应(polymerase chain reaction, PCR)、限制性片段长度多态性(restriction fragment length polymorphism, RFLP)和DNA测序等技术,在正常男性和无精症及少精症不育患者中,对DNMT1基因的SNP rs16999593,rs2228612, rs2228611, KIT基因的SNPrs3819392,以及KITLG基因的SNP rs995030, rs4474514的等位基因频率和基因型频率的分布进行了研究和分析,初步了解这三个基因的变异与生精障碍的关系,为今后进一步阐明这些基因在生精障碍发病过程中的机制提供一些参考依据。 具体研究结如下： 1.通过比较病例组与正常对照组间DNMT1基因SNP rs16999593, rs2228612,rs2228611的等位基因频率和基因型频率的分布,发现SNP rs16999593和rs2228611的等位基因和基因型频率在少精症患者和正常对照组间存在显著性差异。SNP rs16999593的等位基因A(83.7%vs.77.6%, P=0.029, OR=1.47,95%CI=1.044-2.071)和基因型AA(69.1%vs.59%, P=0.027, OR=1.55,95%CI=1.045-2.314)以及SNP rs2228611的基因型AA(18.4%vs.9.9%,P=0.011,OR=2.05,95%CI=1.172-3.586)在少精症患者中显著高于正常对照组。这些结果表明,SNP rs16999593等位基因A和基因型AA,以及SNP rs2228611的基因型AA与少精症的发病风险相关,提示DNMT1基因的多态性可能影响少精症的发病易感性。 2.对KIT基因的SNP rs3819392, KITLG基因的SNP rs995030和rs4474514的等位基因频率和基因型频率分布的研究结果显示,KIT基因SNP rs3819392和KITLG基因SNP rs4474514这两个位点的等位基因频率或基因型频率在少精患者和正常对照组间存在显著性差异。KIT基因的SNP rs3819392的等位基因G(94.2%vs.90.0%, p=0.022)和基因型GG(89.2%vs.82.0%, p=0.042)在少精症患者中显著高于对照组；KITLG基因SNP rs4474514少精症患者基因型CC频率(8.2%vs.3.4%,p=0.034)也显著高于对照组。这些结果表明KIT基因SNPrs3819392和KITLG基因SNP rs4474514的多态性可能与少精症相关。
[Abstract]:Male infertility is a multi-gene genetic disease caused by many factors such as heredity, environment, diet and so on. In recent years, with the increasing incidence of infertility, male infertility has become one of the most common diseases in the world. In the population, about 10 to 15 per cent of the couples are not able to bear normal fertility, of which about half of them are male infertility. The survey shows that approximately 4% to 5% of the male population is male infertility due to genetic factors. Spermatogenesis is one of the most common causes of male infertility, and the genetic factors play an important role in the process of spermatogenesis. In recent years, some progress has been made in the study of the genetic disease of spermatogenic disorder, and some genetic factors related to the spermatogenic disorder, such as the abnormality of the chromosome, the microdeletion of the Y chromosome and the mutation of some genes, have been found. But these factors can only be explained in some cases, and there are still many genetic diseases of spermatogenic disorder that need to be further clarified. Spermatogenesis is a complex process, including the proliferation and differentiation of spermatogonia, meiosis, and the differentiation of sperm cells into sperm. There are a large number of genes involved in this process, and the abnormality of these genes and their expression can lead to the occurrence of spermatogenic disorders. Therefore, the genes involved in the spermatogenic process are also considered as important candidate genes of spermatogenic disorder, and the relationship between these candidate genes and the spermatogenic disorder can be studied, which can provide valuable data for the final elucidation of the pathogenesis of male infertility. DNMT1 (DNMT1) gene is an important candidate gene which has been found in recent years, its coding product DNMT1 is a kind of DNA methylase, which plays an important role in the process of DNA methylation. The study shows that the gene expression regulation is involved in the process of spermatogenesis, and the DNA methylation is an important part of gene expression regulation in the process of spermatogenesis. expression. DNA methylation of certain genes is necessary for normal spermatogenic processes In the process of spermatogenesis, the DNMT1 gene is expressed in the stages of cell proliferation, meiosis, and fine cell differentiation and the mature sperm, and is mainly responsible for catalyzing and maintaining the DNA methylation state after the chromosomal replication, indicating that the DNMT1 gene may play an important role in the normal spermatogenic process. In mice, the abnormal expression of DNMT1 gene can lead to germ cell apoptosis and spermatogenesis. The expression of DNMT1 gene was also found in some patients with spermatogenic disorder. Therefore, the DNMT1 gene may be related to the human spermatogenic disorder, and the genetic variation of the DNMT1 gene may play a role in the spermatogenic disorder. In recent years, the relationship between some signal transduction system and spermatogenic disorder is also gradually affected. The KIT/ KITLG signal transduction system is considered an integral part of the normal development and control of germ cells The KIT receptor is a tyrosine kinase receptor and the product of the KITLG (KIT ligand) gene is a specific ligand KI of the KIT receptor. TLG. The signal system is specifically combined with KITLG via KIT receptor The activation of KIT/ KITLG signal transduction system is critical to the proliferation, meiosis, migration and maturation of germ cells in the testis. The different animal model studies have shown that the abnormality of KIT or KITLG gene can lead to spermatogenic and male infertility. The results of these studies suggest that the variation of KIT and KITLG gene may also be related to human spermatogenesis. In the present, there are no DNMT1 gene, KIT gene and KITLG gene mutation and human spermatogenic disorder. In this study, the SNP rs16999593, rs2228612, rs228611, KIT gene of DNMT1 gene, rs2228612, rs228611 and KIT gene were used in the patients with normal male and azoospermia and oligospermia by using polymerase chain reaction (PCR), restriction fragment length polymorphism (RFLP) and DNA sequencing. 19392 and the distribution of the allele frequency and genotype frequency of the SNP rs995030, rs4474514 of the KITLG gene and the distribution of the genotype frequency. The relationship between the variation of these three genes and the spermatogenic disorder was preliminarily studied, and the mechanism of these genes in the pathogenesis of spermatogenic disorder was further clarified. Some references. The results were as follows:1. The allele and genotype frequency of the SNP rs16999593 and rs228611 were found in the patients with oligospermia and the normal control by comparing the allele frequency and the genotype frequency of the DNMT1 gene SNP rs16999593, rs2228612 and rs228611 between the case group and the normal control group. There was a significant difference between groups. The allele A of the SNP rs16999593 (83.7% vs. 77.6%, P = 0.029, OR = 1.47,95% CI = 1.044-2.071) and genotype AA (69.1% vs.59%, P = 0.027, OR = 1.55,95% CI = 1.045-2.314), and the genotype AA of the SNP rs228611 (18.4% vs. 9.9%, P = 0.011, OR = 2.05,95% CI = 1.172-3.586) were among the patients with oligospermia The results showed that the SNP rs16999593 allele A and the genotype AA, and the genotype AA of the SNP rs228611 were related to the risk of oligospermia, suggesting that the polymorphism of the DNMT1 gene may affect 2. The study of the allele frequency and genotype frequency distribution of the SNP rs3819392, the KITLG gene, the SNP rs995030 and the rs4474514 of the KIT gene showed that the allele frequency or the genotype frequency of both the SNP rs3819392 and the KITLG gene SNP rs4474514 of the KIT gene were in less sperm and normal. There was a significant difference in the control group. The allele G (94.2% vs. 90.0%, p = 0.022) and genotype GG (89.2% vs. 82.0%, p = 0.042) of the KIT gene was significantly higher in the patients with oligospermia than in the control group; the genotype CC frequency of the KITLG gene SNP rs4474514 oligospermia (8.2% vs. 3.4%, p = 0.0 34) was also significantly higher in the control group. These results indicate that the KIT gene SNPrs3819392 and the KITLG gene SNP rs4474514
【学位授予单位】：大理学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R698.2
本文编号：2502532
[Abstract]:Male infertility is a multi-gene genetic disease caused by many factors such as heredity, environment, diet and so on. In recent years, with the increasing incidence of infertility, male infertility has become one of the most common diseases in the world. In the population, about 10 to 15 per cent of the couples are not able to bear normal fertility, of which about half of them are male infertility. The survey shows that approximately 4% to 5% of the male population is male infertility due to genetic factors. Spermatogenesis is one of the most common causes of male infertility, and the genetic factors play an important role in the process of spermatogenesis. In recent years, some progress has been made in the study of the genetic disease of spermatogenic disorder, and some genetic factors related to the spermatogenic disorder, such as the abnormality of the chromosome, the microdeletion of the Y chromosome and the mutation of some genes, have been found. But these factors can only be explained in some cases, and there are still many genetic diseases of spermatogenic disorder that need to be further clarified. Spermatogenesis is a complex process, including the proliferation and differentiation of spermatogonia, meiosis, and the differentiation of sperm cells into sperm. There are a large number of genes involved in this process, and the abnormality of these genes and their expression can lead to the occurrence of spermatogenic disorders. Therefore, the genes involved in the spermatogenic process are also considered as important candidate genes of spermatogenic disorder, and the relationship between these candidate genes and the spermatogenic disorder can be studied, which can provide valuable data for the final elucidation of the pathogenesis of male infertility. DNMT1 (DNMT1) gene is an important candidate gene which has been found in recent years, its coding product DNMT1 is a kind of DNA methylase, which plays an important role in the process of DNA methylation. The study shows that the gene expression regulation is involved in the process of spermatogenesis, and the DNA methylation is an important part of gene expression regulation in the process of spermatogenesis. expression. DNA methylation of certain genes is necessary for normal spermatogenic processes In the process of spermatogenesis, the DNMT1 gene is expressed in the stages of cell proliferation, meiosis, and fine cell differentiation and the mature sperm, and is mainly responsible for catalyzing and maintaining the DNA methylation state after the chromosomal replication, indicating that the DNMT1 gene may play an important role in the normal spermatogenic process. In mice, the abnormal expression of DNMT1 gene can lead to germ cell apoptosis and spermatogenesis. The expression of DNMT1 gene was also found in some patients with spermatogenic disorder. Therefore, the DNMT1 gene may be related to the human spermatogenic disorder, and the genetic variation of the DNMT1 gene may play a role in the spermatogenic disorder. In recent years, the relationship between some signal transduction system and spermatogenic disorder is also gradually affected. The KIT/ KITLG signal transduction system is considered an integral part of the normal development and control of germ cells The KIT receptor is a tyrosine kinase receptor and the product of the KITLG (KIT ligand) gene is a specific ligand KI of the KIT receptor. TLG. The signal system is specifically combined with KITLG via KIT receptor The activation of KIT/ KITLG signal transduction system is critical to the proliferation, meiosis, migration and maturation of germ cells in the testis. The different animal model studies have shown that the abnormality of KIT or KITLG gene can lead to spermatogenic and male infertility. The results of these studies suggest that the variation of KIT and KITLG gene may also be related to human spermatogenesis. In the present, there are no DNMT1 gene, KIT gene and KITLG gene mutation and human spermatogenic disorder. In this study, the SNP rs16999593, rs2228612, rs228611, KIT gene of DNMT1 gene, rs2228612, rs228611 and KIT gene were used in the patients with normal male and azoospermia and oligospermia by using polymerase chain reaction (PCR), restriction fragment length polymorphism (RFLP) and DNA sequencing. 19392 and the distribution of the allele frequency and genotype frequency of the SNP rs995030, rs4474514 of the KITLG gene and the distribution of the genotype frequency. The relationship between the variation of these three genes and the spermatogenic disorder was preliminarily studied, and the mechanism of these genes in the pathogenesis of spermatogenic disorder was further clarified. Some references. The results were as follows:1. The allele and genotype frequency of the SNP rs16999593 and rs228611 were found in the patients with oligospermia and the normal control by comparing the allele frequency and the genotype frequency of the DNMT1 gene SNP rs16999593, rs2228612 and rs228611 between the case group and the normal control group. There was a significant difference between groups. The allele A of the SNP rs16999593 (83.7% vs. 77.6%, P = 0.029, OR = 1.47,95% CI = 1.044-2.071) and genotype AA (69.1% vs.59%, P = 0.027, OR = 1.55,95% CI = 1.045-2.314), and the genotype AA of the SNP rs228611 (18.4% vs. 9.9%, P = 0.011, OR = 2.05,95% CI = 1.172-3.586) were among the patients with oligospermia The results showed that the SNP rs16999593 allele A and the genotype AA, and the genotype AA of the SNP rs228611 were related to the risk of oligospermia, suggesting that the polymorphism of the DNMT1 gene may affect 2. The study of the allele frequency and genotype frequency distribution of the SNP rs3819392, the KITLG gene, the SNP rs995030 and the rs4474514 of the KIT gene showed that the allele frequency or the genotype frequency of both the SNP rs3819392 and the KITLG gene SNP rs4474514 of the KIT gene were in less sperm and normal. There was a significant difference in the control group. The allele G (94.2% vs. 90.0%, p = 0.022) and genotype GG (89.2% vs. 82.0%, p = 0.042) of the KIT gene was significantly higher in the patients with oligospermia than in the control group; the genotype CC frequency of the KITLG gene SNP rs4474514 oligospermia (8.2% vs. 3.4%, p = 0.0 34) was also significantly higher in the control group. These results indicate that the KIT gene SNPrs3819392 and the KITLG gene SNP rs4474514
【学位授予单位】：大理学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R698.2
【参考文献】
相关期刊论文 前1条
1 凡时财;张学工;;DNA甲基化的生物信息学研究进展[J];生物化学与生物物理进展;2009年02期
本文编号：2502532
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