肝豆状核变性基因表达产物及基因突变的研究

A study of Wilson's disease gene encoded products and gene mutations

目的 探讨肝豆状核变性 (Wilson's disease,WD)的发病机理。方法 将活检获得的 WD患者肝标本体外分离、培养肝细胞 ,应用 Western印迹法对 WD患者肝细胞 WD蛋白进行检测 ,同时扩增其基因组 DNA并直接测序。结果 3例 WD患者中有 2例出现肝细胞 WD蛋白特异条带密度降低 ,DNA测序发现其中一例患者存在 ATP7B778位点 CGG→ CTG(Arg778L eu)杂合突变及 770位点 CTC→ CTG改变。结论 WD基因在 WD患者肝细胞的蛋白表达存在异常 ,可能与 ATP7B基因突变有关。

Objective To investigate the pathogenesis of Wilson's disease(WD). Methods Hepatocytes were isolated from WD patients' bioptic hepatic samples and cultured in vitro; WD proteins, the gene putative encoded products, were detected by SDS PAGE in conjunction with Western blotting in liver samples of three patients and two controls. Their genomic DNAs were analyzed by means of direct DNA sequencing of WD gene (ATP7B) on exon 8. Results The WD proteins lanes from two WD patients were found to be much weaker than that from the control, from which one WD patient was proven as heterozygote of 778 position CGG→CTG(Arg778Leu) and 770 position CTC→CTG change of ATP7B. Conclusion WD is highly heterogeneous in clinical manifestations and inheritance pattern. Abnormally expressed putative WD proteins in WD patients might be the results of ATP7B mutations, and the study of ATP7B products would help to probe into the pathogenesis of WD.