家族性与散发性帕金森病Parkin基因4号外显子突变的差异性研究

目的: 通过聚合酶链式反应(PCR)、基因测序技术及TDI-FP技术,检测散发性与家族性帕金森病Parkin基因4号外显子突变的差异性,探讨Parkin基因突变在PD发病机制中的作用,同时建立一种新的突变检测方法.方法: 以36例家族性及114例散发性PD患者为实验对象,150名正常健康人为阴性对照,通过PCR扩增含Parkin基因4号外显子的DNA片段,电泳分析初次筛选片段缺失者.基因测序技术对无缺失片段进行克隆、测序,筛选出突变型及野生型.最后应用TDI-FP技术检测R110或TAMRA标记ddNTP的FP值,鉴定Parkin基因4号外显子的基因点突变型.结果: 在36例家族性PD患者中共发现10例4号外显子缺失突变,占27%,点突变有4例,占11%;114例散发性PD患者中仅有2例4号外显子缺失突变,占1%,而点突变则有48例,占42%.150例阴性对照组中无缺失突变,点突变10例,占6%.结论: Parkin基因突变可能是PD发病的重要因素之一,而散发性与家族性帕金森病具有不同的突变类型,可能与其具有不同的发病机制相关联.

Difference study on sporadic Parkinson's disease and familial Parkinson's disease in exon 4 of Parkin gene

AIM: To study the difference of exon 4 mutation of Parkin gene between sporadic Parkinson's disease (SPD) and familial Parkinson's disease (FPD) by PCR, gene sequence detection and TDI-FP technique, to explore the important role of Parkin gene in the pathogenesis of Parkinson's disease, and to establish a new method for genotyping of mutation patterns. METHODS: The experiment group included 36 FPD patients and 114 SPD patients, and the control group consists of 150 healthy subjects. The target DNA fragment of Parkin gene exon 4 was amplified by polymerase chain reaction and the lost fragments were chosen. A mutation type and a wideness type were established by clone and gene sequence detection. The FP value of the R110 or TAMRA-labeled ddNTP was detected by TDI-FP assay to determine the SNP mutation genotype. RESULTS: Of the 36 cases with FPD, target DNA fragment lost was found in 10 cases (27%) and point gene mutation was found in 4 cases (11%). Of the 114 cases with SPD, target DNA fragment lost was found in only 2 (1%) cases, but point gene mutation were found in 48 (42%) cases. However, in the control group, no target DNA fragment lost was found, while point mutation was found in 10 (6%) cases. CONCLUSION: Parkin gene mutation may be one of the key factors in the pathogenesis of Parkinson's disease. Sporadic Parkinson's disease and familial Parkinson's disease have different mutation patterns, which may be related with the different pathogenesis of SPD and FPD.