以肝病为首发表现的Wilson病患者病理及突变位点分析

Pathological and gene mutation features of Wilson's disease with liver disease as initial manifestation

目的对以肝病为首发表现的Wilson病(Wilson’s disease,WD)患者ATP7B基因外显子进行PCR扩增并测序,研究WD患者基因突变的特点。方法研究对象包括20名无亲缘关系的正常对照者和45例WD患者,提取基因组DNA,扩增外显子的部分片段,并对扩增产物进行测序分析。结果健康对照者未见异常,WD组检测到21种突变,包括4种新发突变,其中12号外显子Lys952Arg和16号外显子Val1140Ala突变频率均为30.2%,为突变热点。肝脏病理主要表现为不同程度的肝脏损伤,48.9%有脂肪变性,所有患者均有不同程度的肝纤维化,86.7%铜染色阳性。结论我国以肝病为首发表现的WD患者ATP7B基因突变是以少数几个热点突变为主和其他广泛存在的少见突变为辅为特征。

Objective To amplify the ATP7B gene of Wilson’s disease(WD)patients with liver disease as initial manifestation by PCR and to sequence the amplification product,so as to characterize the possible mutations.Methods From the genomic DNA of 45 patients with WD,10 exons of ATP7B gene were amplified by polymerase chain reaction,and directed sequencing of the amplification products was performed;20 healthy subjects were served as controls to exclude the possibility of the gene mutation discovered to polymorphism.Results No abnormality was found in the control group.Twenty-one different mutations had been found,in which 4 mutations were novel in WD patients.Mutation frequencies of Lys952Arg in exon 12 and Val1140Ala in exon 16 both reached 30.2%in WD,which may be the hotspots of mutation in Chinese population.The major pathological changes in liver were chronic hepatitis at different degrees,48.9%with fatty degeneration,all patients had different degrees of liver fibrosis,and 86.7%with copper staining positive.Conclusion In China,WD seems resulting from a few relatively common and a large number rare mutations.