先天性巨结肠症WNT8b和SHH基因的突变与表达  

Mutation and expression of WNT8b gene and SHH gene in Hirschprung disease

作　　者：高红[1] 张志波[2] 姜中佳[2] 王大佳[2] 黄英[2] 王维林[3] 

机构地区：[1]中国医科大学附属盛京医院卫生部小儿先天畸形重点实验室,沈阳110004 [2]中国医科大学附属盛京医院小儿外科,沈阳110004 [3]沈阳师范大学生化学院生物科学系

出　　处：《中华胃肠外科杂志》2010年第10期758-761,共4页Chinese Journal of Gastrointestinal Surgery

基　　金：基金项目：国家自然科学基金（30772277）

摘　　要：目的 探讨WNT8b和SHH基因突变与中国儿童先天性巨结肠症（HSCR）发病的关系.方法 收集东北地区72例散发性HSCR患儿术前全血标本（病例组）,同时选取72名性别、年龄相匹配的健康儿童外周血作为对照（对照组）.提取上述外周血标本基因组DNA后,应用PCR方法对WNT8b基因第1外显子和SHH基因第1外显子进行基因突变的检测,将突变样品进行自动测序分析.采用荧光实时定量PCR（qRT-PCR）方法检测外周血WNT8b和SHH基因的mRNA水平.结果 病例组72例HSCR患儿的WNT8b基因测序结果显示,13例WNT8b基因编码区发生突变,其中杂合性缺失（A缺失）8例（11.1%）,碱基置换突变5例（6.9%） SHH基因测序结果显示,11例SHH基因编码区发生突变,其中杂合性缺失（A缺失）7例（9.7%）,碱基置换突变4例（5.6%） 从而引发氨基酸的改变 而对照组均未发现上述突变.病例组与对照组外周血WNT8b和SHH mRNA表达水平分别为30.01±1.13、17.33±0.62和28.25±1.27、18.94±0.31,差异有统计学意义（均P＜0.05）.结论 HSCR患儿外周血WNT8b和SHH基因存在突变和异常表达,这两种基因可能与东北地区中国儿童HSCR的发生有关.Objective To investigate the relationship of WNT8b and SHH genes mutation and Hirschsprung disease（HSCR） in Chinese children. Methods Preoperative whole blood preparations in 72 children with sporadic HSCR from northeast China were collected（study group）. Seventy-two healthy children were used as controls（matched for sex and age）. Genomic DNA was obtained from peripheral blood. Exon 1 of WNT8b gene and the exon 1 of SHH gene were analyzed for gene mutation. The mutation products were automatically sequenced. The levels of WNT8b and SHH Mrna were detected by quantitative real-time PCR（Qrt-PCR） in blood samples. Results On sequencing, 13 out of 72 children with HSCR had WNT8b gene mutation in the coding area, including heterozygosity deletion in 8 cases （11.1%） and base replacement in 5（6.9%）. Eleven children with HSCR had SHH gene mutation in the coding area including heterozygosity deletion in 7 cases（9.7%） and base replacement in 4（5.6%）. No mutations in WNT8b and SHH genes were found in the control group. The WNT8b and SHH Mrna levels were different between the study group and the control group（30.01±1.13 vs. 17.33±0.62,and 28.25±1.27 vs. 18.94±0.31,P〈0.05）. Conclusions WNT8b and SHH mutations and abnormal expressions are present in the peripheral blood of children with sporadic HSCR. These two genes may be related to the development of sporadic HSCR in children in the northeastern China.

关 键 词：先天性巨结肠 基因WNT8b 基因 SHH 突变 

分 类 号：R72[医药卫生—儿科]