抽动秽语综合征候选基因区域15q13-q22.3的遗传学研究

发布时间：2018-06-10 05:54

  本文选题：HDC基因 + 抽动秽语综合征　； 参考：《中南大学》2012年硕士论文

【摘要】：目的：检测中国汉族抽动秽语综合征(Tourette syndrome, TS)患者中组氨酸脱羧酶基因(the histidine decarboxylase gene, HDC)的变异类型及变异频率。 方法：本实验采用两步筛选法,首先采用聚合酶链式反应-单链构象多态技术(polymerase chain reaction-single strand conformation polymorphism, PCR-SSCP)初步筛查100例汉族TS患者中(男/女=78/22,平均年龄：10.7±5.7岁；平均发病年龄：8.0±4.7岁)HDC基因的外显子编码区域及侧翼区域。然后在120例TS患者(包括初步筛查的100例)和240例正常对照(无神经障碍家族史)中对发现的3种变异进一步分析其基因型和等位基因频率分布差异。 结果： 1.我们对100例TS患者进行变异检测,总共发现了3种变异。其中CT碱基替换位于1号外显子编码区下游52bp处(IVS1+52CT),它不改变剪接位点。CA碱基替换(c.426CA)位于4号外显子编码区,是新的沉默变异(G1y142G1y)。GA碱基替换(c.1743GA)位于12号外显子编码区第581位氨基酸(Thr581Thr),是新的变异。 2.我们对这3种变异在TS病例组和正常对照组中的基因型及等位基因分布频率进行了比较。结果表明这3种变异在TS患者及正常对照中无明显差异,提示HDC基因可能不是引起中国汉族TS的主要原因。 3.对该3种变异采用四格表资料方法计算相对危险度值(odd ratio, OR),结果分别显示IVS1+52CT的OR值为1.55(95%CI：0.86-2.79)；c.426CA的OR值为1(95%CI：0.09-11.09)；c.1743GA的OR值为0.83(95%CI：0.43-1.61)。 结论： 1.我们总共发现了3种杂合变异(IVS1+52CT,c.426CA和c.1743GA),其中c.426CA和c.1743GA是新的变异。 2.HDC基因可能不是引起中国汉族TS的主要原因。 目的：探讨染色体15q13-q22.3区域内的基因表达是否与TS相关,鉴定外周血中TS生物标记。 方法：提取外周血淋巴细胞mRNA,逆转录成cDNA,采用实时定量PCR(quantitative realtime PCR, qRT-PCR)初步检测位于染色体15q13-q22.3区域内的7个基因在30例TS患者及30例正常对照人群外周血淋巴细胞中的表达水平差异,7个基因为：组氨酸脱羧酶基因(the histidine decarboxylase gene, HDC), HERC域和类RCC-1域1基因(the HERC domain and RCC-1like domain1gene, HERC1), HERC域和类RCC-1域2基因(the HERC domain and RCC-1like domain2gene, HERC2),胆碱能受体神经烟碱α7基因(the cholinergic receptor nicotinic alpha7gene, CHRNA7),泛素蛋白连接酶E3A基因(the ubiquitin protein ligase E3A gene, UBE3A),泛素特异性肽酶3基因(the ubiquitin specific peptidase3gene, USP3)和淀粉样蛋白前体蛋白结合A2基因(the amyloid precursor protein-binding protein A2gene, APBA2)。结果只有.APBA2基因在TS汉族患者和正常对照人群外周血淋巴细胞中的表达水平有差异(相对表达值为0.19,P0.01),进一步扩大样本,在84例TS患者与100例正常对照组中外周血淋巴细胞中检测APBA2基因的表达水平。 结果： 1.初步在30例TS病例组(男/女=18/12,平均年龄：10.0±3.3岁)及30例正常对照组中(男/女=18/12,平均年龄：11.3±3.8岁)检测7个基因(HDC, HERC1, HERC2, CHRNA7, UBE3A, USP3和APBA2)在外周血淋巴细胞中的mRNA水平,只有APBA2基因存在表达差异(P0.01)。其它6个基因在TS病例组及正常对照组不存在统计学差异(P0.05)。 2.进一步在TS病例组84例(男/女=68/16,平均年龄：9.9±4.0岁)和正常对照组100例(男/女=80/20,平均年龄：10.9±5.9岁)中的分析表明APBA2基因在TS病例组较正常对照组外周血淋巴细胞中的表达下降(相对表达比为0.21,P0.01)。 结论： 1.APBA2基因在中国汉族TS病例组及正常对照组外周血淋巴细胞中存在表达差异(相对表达比为0.21,P0.01),可考虑作为TS潜在的外周血生物标记。 2.基因HDC、HERC1、HERC2、CHRNA7、UBE3A和USP3在TS病例组和正常对照组外周血淋巴细胞中不存在表达差异(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 卤 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 卤 4.7 years old ( mean age : 8.0 卤 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 .

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本文编号：2002197