天津医药 ›› 2015, Vol. 43 ›› Issue (7): 756-760.doi: 10.11958/j.issn.0253-9896.2015.07.015

• 临床研究 • 上一篇    下一篇

RET 基因甲基化在先天性巨结肠发生机制中的作用

张树建, 张辉, 詹江华   

  1. 天津市儿童医院外科


  • 收稿日期:2014-10-14 修回日期:2015-02-10 出版日期:2015-07-15 发布日期:2015-07-15
  • 通讯作者: 詹江华 E-mail:zhanjianghuatj@163.com
  • 基金资助:
    天津市卫生局重点项目

The role of RET proto-oncogene methylation in the pathogenesis of Hirschsprung's disease

ZHANG Shujian, ZHANG Hui, ZHAN Jianghua   

  1. Department of Surgery, Tianjin Childrens Hospital, Tianjin 300074, China

  • Received:2014-10-14 Revised:2015-02-10 Published:2015-07-15 Online:2015-07-15
  • Contact: ZHAN Jianghua E-mail:zhanjianghuatj@163.com

摘要: 目的 研究RET基因(RET proto-oncogene, RET)的甲基化与先天性巨结肠(Hirschsprung’s Disease, HD)发生关系,揭示其在肠壁神经节细胞发生发展中的重要意义。方法 选取2010年1月至2012年3月在天津市儿童医院诊治的先天性巨结肠手术切除病变组织标本,均为扩张段,21例为实验组;同期非HD患儿正常结肠组织标本5例,为阴性对照组。应用亚硫酸氢盐测序法(Bisulfite sequencing PCR,BSP)-直接检测方法检测RET的CpG岛甲基化状态,用免疫组织化学方法检测实验组和对照组中RET蛋白的表达情况。结果 21例实验组标本BSP直接测序方法检测,有12例发现甲基化,甲基化率为12/21(57.14286%);5例阴性对照组标本均未发现甲基化。免疫组化检测21例HD组织标本定量分析:12例甲基化RET的蛋白表达的平均光密度为(0.2009406±0.01474648),另外9例非甲基化RET的蛋白表达的平均光密度为(0.3642093±0.02340409),两组差异具有统计学意义((P<0.05)。结论 RET的CpG岛甲基化会使其蛋白表达水平降低,由此推论RET基因甲基化可能通过影响RET蛋白的表达水平,进而影响到神经节细胞的发育,从而参与HD发生。

Abstract: Abstract: Objective To investigate the relationship between the methylation of RET (proto-oncogene, RET) and
Hirschsprung disease (HD), and understand its significance in the development of intestinal wall ganglion cells. Methods
Twenty-one surgical removal specimens, which were all dilation segment of HD in Tianjin Childrens Hospital, were used as
experimental group, and 5 samples of non-HD normal colon tissues were used as control group. The bisulfite sequencing
(BSP) - direct detecting method was used to detect RET CpG island methylation status. The expression of RET protein was