Wilson病ATP7B基因启动子区突变对基因表达的影响

Effect of the mutation of promoter region in Wilson disease ATP7B gene on the expression of reporter gene

目的探讨Wilson病ATP7B基因启动子区突变与Wilson发病的关系。方法在48例患者中发现2例存在-183(C→T)突变,对发现存在突变的样本的DNA片段进行分离,克隆入pGL2荧光素酶报告基因,转染细胞,测定荧光素酶的活性,以野生型的ATP7B基因启动子作为对照,分析ATP7B基因启动子点突变对转录活性的影响。结果含正常与含突变ATP7B启动子区序列的pGL2载体的荧光素酶转录活性相比较差异无统计学意义(n=3,P>0.05)。结论启动子区-183(C→T)突变不影响基因转录活性。提示该突变的意义有待进一步的探讨。

Objective To find out the relationship between mutation of ATP7B gene promoter region and pathogenesis of Wilson disease（WD）. Methods Two of 48 WD patients presented C→T base substitution mutations at the position - 183. DNA sequences of the promoter region from normal and mutant samples were separated. The fragments containing the promoter region were cloned upstream of the luciferase. Luciferase activity was analyzed. Results The lueiferase activity of reporter gene containing normal sequence of ATP7B gene promoter region did not show significant difference as compared with that of reporter gene containing mutant promoter（ n = 3, P 〉 0. 05）. Conclusion No influence of C→T base substitution mutations on the activity of promoter was observed in study. The results suggest that WD pathogenesis relates little to the mutations of the promoter region in Chinese.