本文選題：SPD + HOXD13��； 參考：《吉林大學(xué)》2012年碩士論文

【摘要】：背景及目的： 先天性并指(趾)多(趾)畸形（SPD）是常見的常染色體顯性遺傳疾病。典型的SPD表現(xiàn)包括3、4手指和4、5足趾并指（趾），伴有完全或不完全的并指(趾)內(nèi)的指(趾)的重復(fù)，該畸形常表現(xiàn)為外顯率不全和表現(xiàn)度的多樣化。HOXD13基因是公認(rèn)的可導(dǎo)致先天性并多指畸形的重要的候選基因。在人類中，HOX基因被認(rèn)為編碼一個(gè)高度保守的轉(zhuǎn)錄因子家族，對(duì)胚胎發(fā)育起重要的作用。HOX基因簇分布在7、17、12、2號(hào)染色體的特定區(qū)域上，位于第2號(hào)染色體HOXD基因簇5’末端的是HOXD13基因，它包含兩個(gè)外顯子，第1外顯子區(qū)域易導(dǎo)致多聚丙氨酸鏈延展突變，第2外顯子區(qū)域易導(dǎo)致堿基缺失及錯(cuò)義突變、無(wú)義突變。EPHA7是比較明確的受HOXD13基因調(diào)節(jié)的下游基因之一,在胚胎組織廣泛表達(dá)，調(diào)節(jié)神經(jīng)系統(tǒng)的發(fā)育、組織分化等多個(gè)方面, HOXD13基因通過(guò)結(jié)合啟動(dòng)子從而調(diào)節(jié)EPHA7下游基因的轉(zhuǎn)錄表達(dá)。 本論文通過(guò)對(duì)中國(guó)一先天性并指(趾)多(趾)畸形（SPD）家系的臨床表型探索及分子遺傳學(xué)研究，進(jìn)一步明確先天性并指畸形的分型，分析可能導(dǎo)致其異常畸形的發(fā)病原因，探討致病基因在胚胎發(fā)育過(guò)程中的作用機(jī)制。 方法： 對(duì)中國(guó)吉林省一例四代并多指(趾)畸形家系進(jìn)行調(diào)查，總結(jié)其家系患者的臨床表型，繪制家系圖譜，分析遺傳學(xué)特征，同時(shí)收集家系成員外周靜脈血，，提取靜脈血DNA，設(shè)計(jì)HOXD13外顯子引物，PCR擴(kuò)增目的基因，將擴(kuò)增后產(chǎn)物直接測(cè)序，分析有無(wú)突變。同時(shí)針對(duì)EPHA7下游基因采用雙熒光素酶報(bào)告基因檢測(cè)對(duì)新發(fā)現(xiàn)的基因突變做進(jìn)一步的功能實(shí)驗(yàn)研究。 結(jié)果： 通過(guò)分析家系圖譜，該患者符合常染色體顯性遺傳。總結(jié)該家系患者臨床表現(xiàn)，符合先天性并指多指畸形特點(diǎn)。通過(guò)分子遺傳學(xué)研究，在本家系所有患者中均發(fā)現(xiàn)HOXD13基因第2外顯子區(qū)域發(fā)現(xiàn)一個(gè)新的錯(cuò)義突變：c.893GA (p.R298Q),即編碼區(qū)第893位堿基由鳥嘌呤（G）突變?yōu)橄汆堰剩ˋ），使得HOXD13基因第298位氨基酸（同源盒結(jié)構(gòu)域區(qū)域第31位）由CGG突變?yōu)镃AG（R31Q），導(dǎo)致精氨酸被谷氨酰胺替代。同時(shí)在家系一例臨床表現(xiàn)正常的患者中發(fā)現(xiàn)同樣的基因突變，顯示其為隱性攜帶者，提示該家系的外顯率不全。我們同時(shí)對(duì)突變體進(jìn)行雙熒光酶報(bào)告基因功能實(shí)驗(yàn)，結(jié)果顯示對(duì)比野生型HOXD13基因，突變型HOXD13（R31Q）對(duì)下游EPHA7基因啟動(dòng)子的轉(zhuǎn)錄活性明顯降低。結(jié)論： 1.本家系顯示出外顯率不全和并指畸形表現(xiàn)度的多樣化，豐富了SPD的臨床表型。 2.本家系研究新發(fā)現(xiàn)的HOXD13基因第2外顯子的錯(cuò)義突變，導(dǎo)致氨基酸的改變，從而影響HOXD13與DNA的結(jié)合，引起HOXD13對(duì)下游基因的轉(zhuǎn)錄結(jié)合能力減弱，推測(cè)該突變可能是導(dǎo)致并指多指畸形的分子機(jī)制之一。
[Abstract]:Background and purpose: Congenital syndactyly polydeformity (SPD) is a common autosomal dominant genetic disease. Typical SPD findings include 3 fingers with 4 fingers and 4 with 5 toes and fingers with complete or incomplete duplication of the fingers (toes) within the fingers (toes). HOXD13 gene is recognized as an important candidate gene for congenital polydactyly deformity. In humans, the HOX gene is thought to encode a highly conserved family of transcription factors that play an important role in embryonic development. HOXD13 gene is located at the 5'end of HOXD gene cluster on chromosome 2. It contains two exons. The region of exon 1 easily leads to polyalanine chain extension mutation, and exon 2 region easily leads to base deletion and missense mutation. Nonsense mutation. EPHA7 is one of the downstream genes regulated by HOXD13 gene. It is widely expressed in embryonic tissues and regulates the development of nervous system. In tissue differentiation and other aspects, HOXD13 gene regulates the transcription and expression of downstream EPHA7 gene by binding to promoter. In this paper, the clinical phenotype and molecular genetics of a Chinese family with congenital syndactyly polydactyly (SPD) were investigated, and the types of congenital syndactyly were further determined, and the possible causes of the abnormal deformity were analyzed. To explore the mechanism of pathogenicity gene in embryonic development. Methods: A family of four generations with polydigital deformity was investigated in Jilin Province, China. The clinical phenotypes of the patients were summarized, family maps were drawn, genetic characteristics were analyzed, and peripheral venous blood was collected at the same time. HOXD13 exon primer was designed to amplify the target gene. The amplified product was sequenced directly and the mutation was analyzed. At the same time, double luciferase reporter gene detection for downstream EPHA7 gene was used to further study the new gene mutation. Results: The pedigree map showed that the patient was autosomal dominant inheritance. The clinical manifestations of this family are conformed to congenital syndactyly deformity. Through molecular genetic research, A new missense mutation in exon 2 of HOXD13 gene was found in all patients of this family. A new missense mutation was found in the region of exon 2 of the HOXD13 gene. A new missense mutation was found, namely, the 893 base of the coding region mutated from guanine G) to adenine An, resulting in the 298th amino acid (homology box) of the HOXD13 gene. The region of domain 31) mutated from CGG to CAGN R31QN, resulting in arginine being replaced by glutamine. At the same time, the same gene mutation was found in a family of patients with normal clinical manifestations, indicating that it was a recessive carrier, indicating that the outlier rate of the pedigree was not complete. At the same time, we performed a double fluorescence reporter gene function experiment on the mutant. The results showed that the transcriptional activity of the mutant HOXD13 / R31Q to the downstream EPHA7 gene promoter was significantly lower than that of the wild type HOXD13 gene. Conclusion: 1. The clinical phenotype of SPD was enriched by the diversity of phenotype of syndactyly. 2. The missense mutation in exon 2 of HOXD13 gene was found in our family, which resulted in the change of amino acid, which affected the binding of HOXD13 to DNA and weakened the transcriptional binding ability of HOXD13 to downstream gene. It is speculated that this mutation may be one of the molecular mechanisms leading to polydactyly deformity.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R726.8

【共引文獻(xiàn)】

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1 楊國(guó)志;李振武;李志安;;Ⅳ型復(fù)拇指初次術(shù)后畸形的矯治[J];中國(guó)醫(yī)藥科學(xué);2013年06期

2 馬會(huì)旭;劉敏;;先天性尺橈骨融合的外科治療進(jìn)展[J];華西醫(yī)學(xué);2012年09期

3 吳西津;趙俊會(huì);;先天性拇指偏斜的手術(shù)治療[J];內(nèi)蒙古醫(yī)學(xué)雜志;2010年S2期

4 周蓉蓉;陳力;李華;;巨指(趾)癥的臨床治療現(xiàn)狀[J];全科醫(yī)學(xué)臨床與教育;2008年02期

5 繆博;王增濤;;指蹼皮瓣滑移術(shù)治療先天性不完全性皮膚性并指畸形[J];山東醫(yī)藥;2006年18期

6 孟光強(qiáng);;手外科修整術(shù)重建手的外形及功能[J];臨床和實(shí)驗(yàn)醫(yī)學(xué)雜志;2010年04期

7 王欣;何漫;章偉文;陳宏;魏鵬;徐吉海;;雙手雙足先天性多指并指畸形(Ⅱ型)1例[J];實(shí)用手外科雜志;2008年02期

8 張愛(ài)麗;陳淑琴;;先天性復(fù)拇畸形的分類及圍手術(shù)期護(hù)理[J];實(shí)用醫(yī)藥雜志;2012年11期

9 湯海萍;滕國(guó)棟;陳艷清;方光榮;孫明川;;遺傳性手關(guān)節(jié)攣縮癥的手術(shù)治療[J];實(shí)用醫(yī)藥雜志;2013年11期

10 劉淑文;郭忠輝;劉蕾;宮立程;楊光;楊軍;;一個(gè)獨(dú)特臨床表型的SPD家系HOXD13基因突變分析[J];中國(guó)實(shí)驗(yàn)診斷學(xué);2012年10期

相關(guān)碩士學(xué)位論文 前4條

1 吳廣智;先天性分裂手畸形的分型與治療[D];吉林大學(xué);2006年

2 吳迪;先天性并指畸形治療的回顧性研究[D];吉林大學(xué);2013年

3 郭志強(qiáng);一先天性并指合并裂手畸形家系致病基因的初步分析[D];吉林大學(xué);2013年

4 劉飛;先天性復(fù)拇指畸形的臨床回顧性研究[D];吉林大學(xué);2014年

本文編號(hào)：1960417