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  本文選題：先天性巨結(jié)腸癥 + RET基因�。� 參考：《山西醫(yī)科大學(xué)》2012年碩士論文

【摘要】：背景 先天性巨結(jié)腸癥（Hirschsprung disease，HSCR）又稱腸無神經(jīng)節(jié)細(xì)胞癥（aganglionosis），是典型的腸神經(jīng)系統(tǒng)發(fā)育異常疾病。其主要病理特征為結(jié)腸或者直腸的肌層及粘膜下層神經(jīng)節(jié)細(xì)胞缺失，腸道神經(jīng)調(diào)節(jié)紊亂，以致受累腸段痙攣收縮，其近端結(jié)腸代償性擴(kuò)張與肥厚，形成巨結(jié)腸，從而導(dǎo)致小兒嚴(yán)重的腸梗阻及便秘等消化道癥狀。 目前的研究顯示多種基因與HSCR發(fā)病有關(guān)，主要有來自腸神經(jīng)系統(tǒng)發(fā)育中起主要作用的2個信號轉(zhuǎn)導(dǎo)通路，RET信號通路和內(nèi)皮素B信號通路。迄今已發(fā)現(xiàn)與之相關(guān)的基因主要有RET原癌基因（RET proto-oncogen）及內(nèi)皮素受體B基因（Endothelin receptor B，EDNRB）。研究學(xué)者推測基因單核苷酸多態(tài)性（single nucleotide polymorphism，SNP）和單倍型可能作為修飾因子存在，增加HSCR患病風(fēng)險。由于遺傳背景等方面的差異，不同的人人群和種族中多態(tài)性存在不同。 目的 研究山西漢族人群先天性巨結(jié)腸癥患兒RET基因和EDNRB基因常見單核苷酸多態(tài)位點與HSCR發(fā)病的關(guān)系。 方法 收集2008年1月至2010年12月在山西省兒童醫(yī)院就診，經(jīng)術(shù)后病理切片證實的80例漢族先天性巨結(jié)腸癥患兒，且均為散發(fā)性。收集同期山西省兒童醫(yī)院體檢健康兒童80例。經(jīng)家長及患兒同意后均取靜脈血3mL，EDTA抗凝。應(yīng)用高分辨率熔解曲線技術(shù)（HighResolution Melt，HRM）以及PCR產(chǎn)物測序，序列比對的方法，，對山西省80例散發(fā)性先天性巨結(jié)腸癥患兒和80例健康兒童進(jìn)行RET基因常見多態(tài)位點p.A45A，p.V125V，p.A432A，p.G691S，p.L769L，p.S904S以及EDNRB基因1-4號外顯子多態(tài)位點的分析。 結(jié)果 1、本課題首次應(yīng)用高分辨熔解曲線技術(shù)并且首次將該技術(shù)對山西漢族先天性巨結(jié)腸癥患兒RET基因單核苷酸多態(tài)性進(jìn)行分析，實驗結(jié)果如下： RET基因p.A45A位點存在多態(tài)性，病例組與對照組相比兩組間等位基因差異顯著（x~2=35.60，P0.01）。p.V125V位點可能不存在基因多態(tài)性。p.A432A位點存在多態(tài)性，病例組和對照組中該位點的等位基因分布頻率存在顯著性差異（x~2=5.668，P=0.017）。p.G691S病例組與對照組兩組比較無明顯差異（x~2=0.7810，P=0.377）。p.L769L位點存在多態(tài)性，病例組和對照組中該位點的等位基因分布頻率存在顯著差異（x~2=37.458，P0.01）。p.S904S存在多態(tài)位點，該位點與疾病無顯著相關(guān)性（x~2=1.053，P=0.305）。 2、EDNRB基因中在第4外顯子發(fā)現(xiàn)多態(tài)位點c831GA，引起亮氨酸的同義突變（p.L277L）。其中A等位基因頻率為0.55，G等位基因頻率為0.45。未發(fā)現(xiàn)其他已報道突變及多態(tài)位點。 結(jié)論 1、在山西漢族人群中，RET基因2號外顯子p.A45A，7號外顯子p.A432A，13號外顯子p.L769L的基因多態(tài)性可能與該地區(qū)先天性巨結(jié)腸癥的發(fā)生密切相關(guān)（P0.05），11號外顯子p.G691S，15號外顯子p.S904S在病例組和對照組比較中無明顯差異（P0.05），在山西漢族人群中，未發(fā)現(xiàn)3號外顯子p.V125V多態(tài)位點。 2、HSCR可檢測到EDNRB基因4號外顯子多態(tài)性改變（p.L277L），在1-4號外顯子中，未發(fā)現(xiàn)其他突變及多態(tài)位點。
[Abstract]:background
Congenital megacolon (Hirschsprung disease, HSCR), also known as intestinal ganglio cell syndrome (aganglionosis), is a typical dysplasia of the intestinal nervous system. Its main pathological features are the absence of ganglion cells in the myometrium and submucosa of the colon or rectum, the disturbance of the intestinal nerve regulation, and the involvement of the intestinal spasm and contraction of the proximal colon. Compensatory dilatation and hypertrophy, forming a megacolon, leading to severe intestinal obstruction and constipation in children.
Current studies have shown that a variety of genes are associated with HSCR, mainly 2 signal transduction pathways, RET signaling pathways and endothelin B signaling pathways, which are mainly derived from the development of the intestinal nervous system. The genes associated with it have been found to be the RET proto oncogene (RET Proto-oncogen) and the endothelin receptor B gene (Endothelin receptor). B, EDNRB). Researchers have speculated that gene single nucleotide polymorphisms (single nucleotide polymorphism, SNP) and haplotypes may exist as modifying factors to increase the risk of HSCR disease. There are different polymorphisms in different human populations and races due to genetic backgrounds.
objective
Objective to investigate the relationship between RET gene and EDNRB gene single nucleotide polymorphisms and HSCR in children with Hirschsprung disease in Shanxi Han population.
Method
From January 2008 to December 2010 in Shanxi children's Hospital, 80 children with Hirschsprung's disease confirmed by postoperative pathological sections were found to be sporadic. 80 healthy children in the medical examination of Shanxi children's hospital were collected for the same period. After the consent of parents and children, the venous blood 3mL and EDTA anticoagulant were taken. The high resolution fusion curve technique was applied. HighResolution Melt (HRM) and the sequencing of PCR products and sequence alignment were carried out in 80 children with sporadic Hirschsprung's disease and 80 healthy children in Shanxi province. The common polymorphic loci of RET gene p.A45A, p.V125V, p.A432A, p.G691S, p.L769L, p.S904S, and the polymorphism of the polymorphic loci of the EDNRB genes were analyzed.
Result
1, the first use of high resolution fusion curve technique and the first analysis of the single nucleotide polymorphisms of RET gene in children with Hirschsprung's disease in Shanxi is the first time. The results are as follows:
The p.A45A locus of RET gene was polymorphic. Compared with the control group, the difference of the allele between the two groups was significant (x~2=35.60, P0.01).P.V125V loci may not exist polymorphism.P.A432A locus, and there was a significant difference in the frequency of allele distribution in the case group and the control group (x~2=5.668, P=0.017).P.G691S disease. There was no significant difference (x~2=0.7810, P=0.377).P.L769L polymorphism between the two groups and the control group. There was a significant difference in the allele distribution frequency of the loci (x~2=37.458, P0.01).P.S904S in the case group and the control group (x~2=37.458, P0.01), and there was no significant correlation between the loci and the disease (x~2=1.053, P=0.305).
2, the polymorphic loci c831GA was found in the fourth exon of EDNRB gene, causing the synonymous mutation of leucine (p.L277L), of which the A allele frequency was 0.55, and the G allele frequency was 0.45. no other reported mutation and polymorphic loci.
conclusion
1, in the Han population of Shanxi, the gene polymorphism of exon 2 of RET gene, exon 7, exon 7 and exon 13 of exon 13 may be closely related to the occurrence of congenital megacolon in this area (P0.05), 11 exon p.G691S, and exon 15 p.S904S in the case group and the control group (P0.05), in the Han nationality of Shanxi. No polymorphisms of exon 3 were found in p.V125V population.
2, HSCR could detect polymorphisms of exon 4 of EDNRB gene (p.L277L), and no other mutations and polymorphic loci were found in exon 1-4.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R726.5

【參考文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

1 郝長鎖;天津地區(qū)先天性巨結(jié)腸RET基因13號外顯子基因多態(tài)性研究[D];天津醫(yī)科大學(xué);2009年

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