RET基因多态性与先天性巨结肠患病风险研究

发布时间：2018-11-12 17:53

【摘要】：目的：先天性巨结肠(Hirschsprung's Disease, HSCR)又称肠管无神经节细胞症,是小儿常见的先天性消化道畸形,发病率居先天性消化道畸形第二位,以不同病变范围内肠段肌间神经丛和粘膜下神经丛神经节细胞的缺失为主要病理学特征。目前研究证明它是一种的多基因型疾病,是多个基因位点相互作用的结果,其中最主要的基因是RET原癌基因(RET proto-oncogen)。RET基因具有多态性和单倍型,可能作为基因修饰因子,而且可能神经嵴胚细胞发育过程中发生紊乱的危险性增加与之相关。本研究通过深圳地区先天性巨结肠患儿RET基因上intronlCT (rs2435357)单核苷酸多态性的研究及基因多态表达情况,明确其与HSCR的关联性,揭示intronlCT(rs2435357)多态性在深圳地区人群散发性HSCR发病中所起的重要作用,探讨其与HSCR发病风险的关系,并为HSCR的发病机制提供线索。方法：收集2008年5月至2011年8月在深圳市儿童医院外科诊治的HSCR患儿的静脉全血标本115例,均为散发性,其中男90例,女25例,患有TCA(全结肠型先天性巨结肠),L-HSCR(长段型先天性巨结肠)和S-HSCR(短段型先天性巨结肠)的病人分别为4人,6人和105人,年龄17天-81天,无先天性巨结肠家族性遗传病史。所有患者均经术后病理组织学确认为HSCR。收集139名性别和年龄与之匹配的来源于深圳市儿童医院体检的正常儿童静脉全血标本作为对照组。抽取研究样本静脉全血3ml使用QIAamp-Blood Kit (Qiagen, Hilden,Germany)试剂盒进行提取血样本基因组DNA,聚合酶链反应(polymerase chain reaction, PCR)扩增RET基因intronl CT(rs2435357),然后应用直接测序方法分析intronl CT(rs2435357)的基因多态性改变,对基因多态性位点等位基因频率在两组中的差异进行比较,比较不同基因型与先天性巨结肠风险的相关性。回归模型中使用OR值和95%置信区间(CI)作为基因型危险性的评价指标。结果：RET基因intronlCT(rs2435357)基因频率在病例和正常对照人群的分布具有显著差异(P0.001),携带RET基因intronlTT基因型者罹患先天性巨结肠的风险是携带RET intron1CC基因型者的19.22倍(95%CI,7.54-48.99)。RET基因intronl TT(OR=19.22,95%CI=7.54-48.99)基因型能明显增加HSCR发病的风险。结论：RET基因intronlCT(rs2435357)单核苷酸多态可能是深圳地区发生先天性巨结肠的遗传易感因素
[Abstract]:Objective: congenital Hirschsprung's disease (Hirschsprung's Disease, HSCR) is a common congenital alimentary tract malformation in children. The main pathological features were the absence of ganglion cells in the myenteric plexus and submucosal plexus in different lesions. At present, it has been proved to be a multi-genotypic disease, which is the result of the interaction of multiple gene loci. The most important gene is the proto-oncogene of RET (RET proto-oncogen). RET gene is polymorphic and haplotype). It may be a genetically modified factor and may be associated with an increased risk of disorder in the development of neural crest embryonic cells. In this study, we studied the single nucleotide polymorphisms of intronlCT (rs2435357) on RET gene and gene polymorphism expression in children with Hirschsprung's disease in Shenzhen area to determine its association with HSCR. To reveal the important role of intronlCT (rs2435357) polymorphism in sporadic HSCR in Shenzhen area, to explore its relationship with the risk of HSCR, and to provide clues for the pathogenesis of HSCR. Methods: from May 2008 to August 2011, the venous blood samples of 115 patients with HSCR were collected, including 90 males and 25 females, with TCA (total colonic Hirschsprung's disease). Patients with L-HSCR (long segment Hirschsprung's disease) and S-HSCR (short segment Hirschsprung's disease) were 4, 6 and 105, respectively. Their age was 17 days to 81 days. There was no history of familial hereditary disease of Hirschsprung's disease. All patients were confirmed as HSCR. by postoperative histopathology. A total blood sample of 139 healthy children with matched sex and age was collected from Shenzhen Children's Hospital as control group. The genomic DNA, polymerase chain reaction (polymerase chain reaction, PCR) amplification of RET gene intronl CT (rs2435357 was carried out by using QIAamp-Blood Kit (Qiagen, Hilden,Germany kit to extract the whole blood 3ml from the study samples. Then the genetic polymorphisms of intronl CT (rs2435357) were analyzed by direct sequencing. The allelic frequencies of polymorphic loci were compared between the two groups, and the correlation between different genotypes and the risk of Hirschsprung's disease (Hirschsprung's disease) was compared. OR value and 95% confidence interval (CI) were used to evaluate genotype risk in regression model. Results: the frequency of intronlCT (rs2435357) gene of RET gene was significantly different between the case and the control group (P0. 001). People with intronlTT genotype of RET gene were 19.22 times more likely to develop congenital megacolon than those with RET intron1CC genotype. The 7.54-48.99). RET gene intronl TT (OR=19.22,95%CI=7.54-48.99) genotype significantly increased the risk of HSCR. Conclusion: the single nucleotide polymorphism of RET intronlCT (rs2435357) may be the genetic susceptibility factor of Hirschsprung's disease in Shenzhen area.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R726.5

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