Objective To study the expression of ATP7BcDNA in the the hepatocyte of Wilson patients in Vitro,and obersve the expression of ATP7B protein in various periods. Methods Hepatocyte from Wilson patients were isolated and cultured in vitro,target gene ATP7BcDNA was transfect into the hepatocyte through viral vectors,the expression of ATP7B protein in various periods by immunocytochemistry and enzyme linked immunosorbent assay. Results The synthesis of ATP7B protein in the hepatocytes was confirmed by immunocytoehemistryc (ICC ) ,the expression of ATP7B protein began to increase at 3h,peaked at 3-5d,then decreased gradually after 15days. Conclusion Synthesis of ATP7B protein was confirmed successfully when restructuring adenovirus vector with ATP7B were trans-fected into the cutured hepatocyte of Wilson patients.%目的 研究腺病毒介导的ATP7B基因转染肝豆状核变性(Wilson)患者的肝细胞后,ATP7B在肝细胞内的表达及生物学活性.方法 ATP7B基因重组腺病毒载体转染体外培养的Wilson患者肝细胞,观测绿色荧光在肝细胞内的表达,以细胞免疫化学技术和ELISA等方法检测ATP7B蛋白在细胞内、外的表达情况.结果 经转染后肝细胞能成功合成ATP7B蛋白,其表达量在转染后的3～5d达到高峰,14 d后表达显著下降.结论 携带ATP7B目的基因的重组腺病毒载体经转染后能在Wilson病离体肝细胞中成功表达.
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