DVL-1和DVL-3基因在先天性巨结肠症肠管中的表达  

The Expression of dishevelled-1 and dishevelled-3 genes in colons of Hirschsprung＇s disease

作　　者：陈东[1] 弭杰[1] 伍美[1] 高红[1] 

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

出　　处：《中华小儿外科杂志》2014年第1期51-55,共5页Chinese Journal of Pediatric Surgery

基　　金：国家自然科学基金资助项目（30772277）

摘　　要：目的通过dishevelled-1（DVL-1）和dishevelled-3（DVL-3）基因及其相关蛋白在先天性巨结肠症（Hirschsprung＇s disease，HD）中表达水平的研究，探索DVDl和DVD3与HD的发病关系。方法利用荧光实时定量PCR（quantitative real-time polymerase chain reaction, qRT-PCR）、蛋白质印迹（Westernblot）和免疫组织化学方法检测50例HD患儿狭窄段（无神经节细胞）和正常段（有神经节细胞）肠管组织DVL-1和DVL-3的表达情况，并对其表达进行定量分析与比较。结果DV卜1在HD狭窄段肠管中mRNA表达量是正常段肠管中的2．06倍；DVD3在HD狭窄段肠管中mRNA表达量是正常段肠管中的3．12倍，差异有统计学意义（P〈0．03）。Westernblot结果显示，DVDl在HD狭窄段肠管中蛋白相对表达量为39．71±4．53，高于正常段肠管蛋白表达量为15．01±2．66，差异有统计学意义（P〈0．05）；DVD3在HD狭窄段肠管中蛋白相对表达量为53．90土6．79，高于正常段肠管蛋白的表达量20．13±3．63，差异有统计学意义（P〈0．05）。免疫组化结果显示，DVDl和DVD3在HD狭窄段肠管的黏膜层细胞胞质内呈强阳性反应，呈深棕黄色，而在HD正常段肠壁的黏膜层细胞胞质内呈阴性或弱阳性反应，无色或淡黄色；此外，研究还发现，与正常段的神经丛相比，狭窄段肠管中异常增生的神经纤维组织内DVL-3的表达信号更强。结论DVD1和DVD3mRNA与蛋白在HD狭窄段肠管中高表达，提示DVD1和DVL-3与HD的发生有密切关系，可能在先天性消化道畸形的肠道发育中具有重要作用；DVD3可能成为今后临床诊断HD一项参考指标。Objective To evaluate the expression of dishevelled-1 （DVL-1） and dishevelled-3 （DVL-3） genes and proteins in the aganglionic and normal segment of colon in patients with Hirschsprung＇s disease （HSCR）. Methods The expression of DVL-1 and DVL-3 genes were detected by quantitative real-time polymerase chain reaction （ qRT-PCR ）, Western blot and immunohistochemical staining in the aganglionic and normal segment of colon in 50 patients with HSCR. Results The mRNA expression of DVL-1 and DVL-3 was 2. 06 fold and 3. 12 fold in the aganglionic segment colon tissues compared to the normal segment, respectively. Similarly, the protein expression of DVL-1 and DVL-3 was higher in the stenotic colon segments than that in the normal segment （39.71 ± 4.53 vs. 15.01 ± 2.66; 53.90 ± 6.79 vs. 20. 13 ± 3.63, respectively）. Besides, immunohistochemical staining showed that the expression of DVL-1 and DVL-3 significantly increased in the mucous and submucous layers from stenotic colon segments compared to the normal segments. Conclusions Higher expression of DVL-1 and DVL-3 mRNA and protein in the stenotic colon of HD suggests that DVL-1 and DVL-3 are involved in the pathogenesis of HE）. DVL-3 may become a reference index for clinical diagnosis of HD in the future.

关 键 词：先天性巨结肠 蓬乱蛋白 聚合酶链反应 

分 类 号：R726.5[医药卫生—儿科]