Expression and clinical significance of Wnt10b gene in Hirschsprung disease / 中华消化外科杂志

ABSTRACT

Objective To study the expression and clinical significance of Wnt10b gene in tissue samples of Hirschsprung disease (HD).Methods Sixty samples of stenotic intestines and 60 samples of normal intestines were collected from 60 patients with sporadic HD who were admitted to the Shengjing Hospital of China Medical University from 2003 to 2010.Tissue samples were stained by hematoxylin and eosin (HE),and then the expression of Wnt10b gene in the tissues was detected by immunohistochemical staining.The mRNA and protein expressions of Wnt10b in HD were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot.All data were analyzed by two independent sample t test.Results The results of HE staining confirmed HD in the 60 patients,and the samples met the requirement of the study.The expressions of Wnt10b were strong positive in the plasma of intramuscular and submucosal cells in stenotic intestines,while weak positive or negative in normal intestines.The results of immunohistochemical staining showed that the percentages of positive proteins in the stenotic intestines and the normal intestines were 0.061％ ± 0.014％ and 0.006％ ± 0.005％,respectively,with a significant difference (t =2.955,P ＜ 0.05).The results of qRT-PCR showed that the relative quantification of mRNA of Wnt10b was 23.5 ± 1.6 in the stenotic intestines,which was significantly higher than 13.1 ± 1.7 in the normal intestines (t =1.687,P ＜0.05).High mRNA expressions of Wnt10b were observed in 45 patients with HD,and low mRNA expressions of Wnt10b were observed in 15 patients.The results of Western blot showed that the relative protein expression of Wnt10b was 35.2 ± 2.3 in the stenotic intestines,which was significantly higher than 19.1 ± 1.3 in the normal intestines (t =2.046,P ＜ 0.05).High protein expressions of Wnt10b were observed in 43 patients,and low protein expressions of Wnt10b were observed in 17 patients.Conclusion There is a close relationship between the abnormal mRNA and protein expressions of Wnt10b in the intestines,which might play a role in the pathogensis congenital malformation of the alimentary tract.