Study Of RET And EDNRB DNA Polymorphism In Patients With Hirschsprung’s Disease In Shanxi Province

BackgroundHirschsprung disease also known as aganglionosis is the typical development disease of theenteric nervous system disordered. It main pathological of the muscularis and submucosa of thecolon or rectum in the ganglion is cell loss, the nervous adjustment disordered. It resulted inbowel spasm contraction and compensatory expansion of the proximal colon. So there will beserver ileus and constipation and other gastrointestinal symptoms in children.The present study shows that many genes are related to HSCR. There are two major signalsfrom the enteric nervous system that play a major role in the transduction pathway, RETsignaling pathway and endothelin signaling pathway. Scholars speculated that single nucleotidepolymorphism (SNP) and haplotypes may be used as a genetic modifier factors, increasing therisk of HSCR. There are difference between ethnic and racial because of difference in geneticand background.ObjectiveThe aim of this study is to analyze single nucleotide polymorphisms in the RETproto-oncogen and endothelin receptor B with the etiopathogenisis of HSCR in ShanXiprovincial patients of Han.MethodsCollected 80 cases of children with Hirschsprung’s disease confirmed by postoperativepathology from January 2008 to December 2010 in the Shanxi Children’s Hospital for treatment.They are sporadic. At the same time, collected 80 healthy children of the Children’s hospital ofShanxi Province. Blooding venous 3ml after the consent of the parents and children. By HighResolution Melt, PCR products sequenced, analysis p.A45A，p.V125V，p.A432A，p.G691S， p.L769L，p.S904S in RET and exon1-4 in EDNRB of the 80 cases of Hirschsprung’s disease and80 healthy children.ResultsIt is the first time to analysis RET and EDNRB gene with the Hirschsprung disease withHigh Resolution Melt in ShanXi provincial patients of Han .The results are as follows:1.There are polymorphism p.A45A in RET gene. There are difference between case groupand control group(x~2=35.60，P<0.01).There may not exist polymorphism in p.V125V. There arepolymorphism p.A432A in RET gene. There are difference between case group and controlgroup(x~2=5.668，P=0.017). There are no significant difference between case group and controlgroup in p.G691S（x~2=0.7810，P=0.377）. There are polymorphism p.L769L in RET gene. Thereare difference between case group and control group（x~2=37.458，P<0.01）There arepolymorphism p.S904S. There are no significant difference between case group and controlgroup（x~2=1.053，P=0.305）.2.There is a polymorphism c831G>A in exon 4 of EDNRB, causing leucine synonymousmutation(p.L277L).The allele frequency of A is 0.55,and G is 0.45.The other reported mutationsand polymorphism are not found.Conclusions1. p.A45A, p.A432A and p.L769L in RET gene are closely related to the occurrence ofHirschsprung disease in Han population in ShanXi（P<0.05）.There are no significant differencebetween case group and control group about p.G691S and p.S904S（P>0.05）.There are nop.V125V in Han population in ShanXi.2. In HSCR, p.L277L can be found in exon 4 of EDNRB ,but the other reported mutationsand polymorphism in exon1-4 are not found.