血管内皮生长因子受体2遗传变异与子宫内膜异位症发病风险的关联研究

发布时间：2018-03-17 07:27

  本文选题：子宫内膜异位症　切入点：血管内皮生长因子受体　出处：《河北医科大学》2014年硕士论文　论文类型：学位论文

【摘要】：目的：子宫内膜异位症（endometriosis，Ems，简称内异症）指具有活性的子宫内膜腺体或间质成分在子宫内膜及子宫肌层以外出现、生长、浸润、反复出血、可形成结节及包块，引起疼痛和不育。子宫内膜异位症是妇科常见疾病，在育龄期妇女其发病率达10%。研究表明：在内异症的发病过程中，内异症病灶中血管生成及血液供应起重要作用。新生血管形成是内异症病变形成中重要的环节，这一观点基本被认可。血管内皮生长因子（Vascular Endothelial Growth Factor，VEGF）是生理性及病理性血管生成最重要调节物，VEGF主要通过结合其酪氨酸激酶受体VEGFR-2发挥其促血管生成生物活性。本研究旨在探讨VEGFR-2基因上的单核苷酸多态性（Single Nucleotide Polymorphisms，SNPs）位点与子宫内膜异位症发病风险的关系，其结果可能从基因水平进一步探讨内异症的发病机制，并为其预防及诊治提供重要理论依据。 方法：本实验采用病例-对照研究，分析VEGFR-2基因上的遗传变异与内异症发病风险的关系。研究对象包括571例内异症患者（病例组），580例健康对照个体（对照组）。采集研究对象的外周静脉血5ml，采用蛋白酶K消化-饱和氯化钠盐析法提取外周血白细胞DNA。根据HapMap数据库中VEGFR-2基因上的中国北方汉族人群的数据，利用Haploview软件，通过htSNPs（haplotype tag Single Nucleotide Polymorphisms）的方法对VEGFR-2基因上的SNP位点进行筛选，最终确定了5个多态性位点：+1192C/T（rs2305948）、+1719T/A（rs1870377）、IVS6+54C/T（rs7692791）、IVS25-92G/A（rs1531289）和+31C/T（rs7667298）。采用聚合酶链反应和连接酶检测反应（polymerase chain reaction-ligase detection reaction，PCR-LDR）技术，检测病例组和对照组单核苷酸多态性位点的基因型和等位基因型频率分布情况。数据统计分析采用SPSS13.0版软件包（SPSSCompany，Chicago，Illinois，USA）进行分析，P0.05被认为差异有统计学意义。病例组和对照组的年龄差异采用t检验。对照组基因型频率分布行χ2检验，进行Hardy-weinberg平衡分析。比较病例组和对照组各位点基因型和等位基因频率分布采用χ2检验。以非条件Logistic回归方法计算相对风险度的比值比（odds ratio，OR）及其95%可信区间（confidenceinterval，CI）。 结果： 1.内异症患者的年龄与其对照组相比，差异无统计学意义（P＞0.05）。统计学分析显示对照组中VEGFR-2基因上的5个多态位点的基因型频率分布均符合Hardy-weinberg平衡（P＞0.05）。 2. VEGFR-2基因+1192C/T多态的C和T等位基因频率在对照组和内异症组分别为：86.0%，14.0%和89.3%，10.7%，两组相比存在明显差异（P=0.017）。与C/C基因型相比，C/T+T/T基因型的携带者内异症的发病风险明显降低，OR值为0.75（95%CI=0.57-0.99）。 3. VEGFR-2基因+1719T/A多态的T/T，T/A，A/A三种基因型频率在对照组和内异症组中分别为：30.2%，50.7%，19.1%和30.3%，49.0%，20.7%，差异无统计学意义（P=0.780）；T和A等位基因频率分别为：55.5%，44.5%和54.8%，45.2%，两组相比无差异（P=0.735）。 4. VEGFR-2基因+31C/T多态的C/C，C/T，T/T三种基因型频率在对照组和内异症组中分别为：41.4%，47.6%，11.0%和42.2%，48.3%，9.5%，差异无统计学意义（P=0.677）；C和T等位基因频率分别为：65.2%，34.8%和66.4%，33.6%，两组相比无差异（P=0.543）。 5. VEGFR-2基因-92G/A多态的G/G，G/A，A/A三种基因型频率在对照组和内异症组中分别为：62.6%，33.8%，3.6%和65.7%，31.0%，，3.3%，差异无统计学意义（P=0.551）；G和A等位基因频率分别为：79.5%，20.5%和81.2%，18.8%，两组相比无差异（P=0.308）。 6. VEGFR-2基因+54T/C多态的T/T，T/C，C/C三种基因型频率在对照组和内异症组中分别为：34.7%，49.7%，15.7%和36.6%，46.4%，17.0%，差异无统计学意义（P=0.540）；T和C等位基因频率分别为：59.5%，40.5%和59.8%，40.2%，两组相比无差异（P=0.955）。 结论： VEGFR-2基因外显子7区+1192C/T多态性位点可能与中国北方汉族妇女子宫内膜异位症的发病风险相关，与C/C基因型相比，C/T+T/T基因型的携带者子宫内膜异位症的发病风险明显降低。VEGFR-2基因+1719T/A，+31C/T，-92G/A，+54C/T多态性位点可能与中国北方汉族妇女子宫内膜异位症的发病风险无关。
[Abstract]:Objective: Endometriosis (endometriosis, Ems, referred to as endometriosis) refers to endometrial glands with active or stromal elements in the endometrium and myometrium outside appearance, growth, invasion, repeated hemorrhage, can form nodules and mass, causing pain and infertility. Endometriosis is a common gynecological disease and in women of childbearing age and the incidence rate of 10%. research shows that: in the pathogenesis of endometriosis and endometriosis lesions in vascular formation and blood supply plays an important role. Neovascularization is an important link in endometriosis lesion formation, this point is recognized. Vascular endothelial growth factor (Vascular Endothelial Growth Factor. VEGF) is the physiological and pathological angiogenesis is the most important regulator of VEGF, mainly by binding to its receptor tyrosine kinase VEGFR-2 plays in promoting the activity of angiogenesis. The purpose of this study was to investigate the biological in VEGFR-2 The relationship between Single Nucleotide Polymorphisms (SNPs) loci and the risk of endometriosis may result from further investigation of the pathogenesis of endometriosis at gene level, and provide important theoretical basis for its prevention and treatment.
Methods: This study used a case-control study, the relationship between the genetic variation analysis of VEGFR-2 gene and the pathogenesis of endometriosis risk. The study included 571 cases of endometriosis patients (case group), 580 healthy individuals (control group). Collected peripheral venous blood 5ml, using proteinase K digestion saturated sodium chloride salting out extraction of peripheral white blood cells of DNA. according to the VEGFR-2 HapMap database on the gene China hanpopulation data, using Haploview software, through htSNPs (haplotype tag Single Nucleotide Polymorphisms) is the method of SNP VEGFR-2 alleles were identified 5 polymorphic loci: +1192C/T (rs2305948) +1719T/A, (rs1870377), IVS6+54C/T (rs7692791), IVS25-92G/A (rs1531289) and +31C/T (rs7667298). Using polymerase chain reaction and ligase detection reaction (polymerase chain reaction-liga Se detection reaction, PCR-LDR) detection technology, the case group and the control group of single nucleotide polymorphism genotype and allele frequency distribution. Using SPSS13.0 software package for statistical analysis of data (SPSSCompany, Chicago, Illinois, USA) analysis, P0.05 was considered statistically significant. The age difference between cases and controls group by t test. The control group genotype frequency distribution for 2 test, Hardy-weinberg analysis. The balance between case group and control group each genotype and allele frequency distribution of 2 test was used. With no conditional Logistic regression models were used to calculate oddsratios (odds ratio, OR) and 95% confidence interval (confidenceinterval, CI).
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