本文選題：HDC基因 + 抽動穢語綜合征��； 參考：《中南大學(xué)》2012年碩士論文

【摘要】：目的：檢測中國漢族抽動穢語綜合征(Tourette syndrome, TS)患者中組氨酸脫羧酶基因(the histidine decarboxylase gene, HDC)的變異類型及變異頻率。 方法：本實驗采用兩步篩選法,首先采用聚合酶鏈?zhǔn)椒磻?yīng)-單鏈構(gòu)象多態(tài)技術(shù)(polymerase chain reaction-single strand conformation polymorphism, PCR-SSCP)初步篩查100例漢族TS患者中(男/女=78/22,平均年齡：10.7±5.7歲；平均發(fā)病年齡：8.0±4.7歲)HDC基因的外顯子編碼區(qū)域及側(cè)翼區(qū)域。然后在120例TS患者(包括初步篩查的100例)和240例正常對照(無神經(jīng)障礙家族史)中對發(fā)現(xiàn)的3種變異進(jìn)一步分析其基因型和等位基因頻率分布差異。 結(jié)果： 1.我們對100例TS患者進(jìn)行變異檢測,總共發(fā)現(xiàn)了3種變異。其中CT堿基替換位于1號外顯子編碼區(qū)下游52bp處(IVS1+52CT),它不改變剪接位點。CA堿基替換(c.426CA)位于4號外顯子編碼區(qū),是新的沉默變異(G1y142G1y)。GA堿基替換(c.1743GA)位于12號外顯子編碼區(qū)第581位氨基酸(Thr581Thr),是新的變異。 2.我們對這3種變異在TS病例組和正常對照組中的基因型及等位基因分布頻率進(jìn)行了比較。結(jié)果表明這3種變異在TS患者及正常對照中無明顯差異,提示HDC基因可能不是引起中國漢族TS的主要原因。 3.對該3種變異采用四格表資料方法計算相對危險度值(odd ratio, OR),結(jié)果分別顯示IVS1+52CT的OR值為1.55(95%CI：0.86-2.79)；c.426CA的OR值為1(95%CI：0.09-11.09)；c.1743GA的OR值為0.83(95%CI：0.43-1.61)。 結(jié)論： 1.我們總共發(fā)現(xiàn)了3種雜合變異(IVS1+52CT,c.426CA和c.1743GA),其中c.426CA和c.1743GA是新的變異。 2.HDC基因可能不是引起中國漢族TS的主要原因。 目的：探討染色體15q13-q22.3區(qū)域內(nèi)的基因表達(dá)是否與TS相關(guān),鑒定外周血中TS生物標(biāo)記。 方法：提取外周血淋巴細(xì)胞mRNA,逆轉(zhuǎn)錄成cDNA,采用實時定量PCR(quantitative realtime PCR, qRT-PCR)初步檢測位于染色體15q13-q22.3區(qū)域內(nèi)的7個基因在30例TS患者及30例正常對照人群外周血淋巴細(xì)胞中的表達(dá)水平差異,7個基因為：組氨酸脫羧酶基因(the histidine decarboxylase gene, HDC), HERC域和類RCC-1域1基因(the HERC domain and RCC-1like domain1gene, HERC1), HERC域和類RCC-1域2基因(the HERC domain and RCC-1like domain2gene, HERC2),膽堿能受體神經(jīng)煙堿α7基因(the cholinergic receptor nicotinic alpha7gene, CHRNA7),泛素蛋白連接酶E3A基因(the ubiquitin protein ligase E3A gene, UBE3A),泛素特異性肽酶3基因(the ubiquitin specific peptidase3gene, USP3)和淀粉樣蛋白前體蛋白結(jié)合A2基因(the amyloid precursor protein-binding protein A2gene, APBA2)。結(jié)果只有.APBA2基因在TS漢族患者和正常對照人群外周血淋巴細(xì)胞中的表達(dá)水平有差異(相對表達(dá)值為0.19,P0.01),進(jìn)一步擴(kuò)大樣本,在84例TS患者與100例正常對照組中外周血淋巴細(xì)胞中檢測APBA2基因的表達(dá)水平。 結(jié)果： 1.初步在30例TS病例組(男/女=18/12,平均年齡：10.0±3.3歲)及30例正常對照組中(男/女=18/12,平均年齡：11.3±3.8歲)檢測7個基因(HDC, HERC1, HERC2, CHRNA7, UBE3A, USP3和APBA2)在外周血淋巴細(xì)胞中的mRNA水平,只有APBA2基因存在表達(dá)差異(P0.01)。其它6個基因在TS病例組及正常對照組不存在統(tǒng)計學(xué)差異(P0.05)。 2.進(jìn)一步在TS病例組84例(男/女=68/16,平均年齡：9.9±4.0歲)和正常對照組100例(男/女=80/20,平均年齡：10.9±5.9歲)中的分析表明APBA2基因在TS病例組較正常對照組外周血淋巴細(xì)胞中的表達(dá)下降(相對表達(dá)比為0.21,P0.01)。 結(jié)論： 1.APBA2基因在中國漢族TS病例組及正常對照組外周血淋巴細(xì)胞中存在表達(dá)差異(相對表達(dá)比為0.21,P0.01),可考慮作為TS潛在的外周血生物標(biāo)記。 2.基因HDC、HERC1、HERC2、CHRNA7、UBE3A和USP3在TS病例組和正常對照組外周血淋巴細(xì)胞中不存在表達(dá)差異(P0.05)。
[Abstract]:Objective : To detect the mutation type and mutation frequency of histidine decarboxylase gene ( HDC ) in Tourette syndrome ( TS ) patients in Chinese Han nationality .

Methods : A two - step method was used to screen 100 Han TS patients ( male / female = 78 / 22 , mean age : 10.7 鹵 5.7 years ) by polymerase chain reaction - single strand conformation polymorphism ( PCR - SSCP ) .
The exon coding region and flanking region of the HDC gene were 8.0 鹵 4.7 years old ( mean age : 8.0 鹵 4.7 years ) . In 120 patients with TS ( 100 patients including primary screening ) and 240 normal controls ( family history without neurological disorder ) , the genotypes and allelic frequency distribution were further analyzed .

Results :

1 . A total of three mutations were found in 100 TS patients . The CT base substitution was located at 52bp downstream of exon 1 ( IVS1 + 52CT ) , which did not alter the splice site . The substitution of CA base ( c.426CA ) was located in exon 4 of exon 4 . The substitution of CA base ( c.1743GA ) was located in exon 1 of exon 12 of exon 12 . The substitution of GA base ( c.1743GA ) was a new mutation .

2 . We compared the genotype and allelic frequency of these three variants in TS case group and normal control group . The results showed that these three variants had no significant difference in TS patients and normal controls , suggesting that HDC gene might not be the main cause of TS in Chinese Hans .

3 . The results showed that the OR value of IVS1 + 52CT was 1.55 ( 95 % CI : 0.86 - 2.79 ) .
c . The OR value of 426CA was 1 ( 95 % CI : 0.09 - 11.09 ) ;
c . The OR value of 1743GA was 0.83 ( 95 % CI : 0.43 - 1.61 ) .

Conclusion :

1 . A total of 3 heterozygous mutations ( IVS1 + 52CT , c.426CA and c.1743GA ) were found in which c . 426CA and c.1743GA were new variants .

2 . HDC gene may not be the main cause of TS in Chinese Hans .

Objective : To investigate whether gene expression in chromosome 15q13 - q22.3 is related to TS and to identify TS biomarkers in peripheral blood .

鏂規(guī)硶錛氭彁鍙栧鍛ㄨ娣嬪反緇嗚優(yōu)mRNA,閫嗚漿褰曟垚cDNA,閲囩敤瀹炴椂瀹氶噺PCR(quantitative realtime PCR, qRT-PCR)鍒濇媯，

本文編號：2002197