| 人肝癌特异性 HSV-TK/GCV重组腺病毒相关病毒质粒的构建与研究 |
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| 引用本文： | 李志辉,严律南,赵永恒,刘智敏,刘自明,李玉. 人肝癌特异性 HSV-TK/GCV重组腺病毒相关病毒质粒的构建与研究[J]. 中国普外基础与临床杂志, 2004, 11(3): 234-237 |
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| 作者姓名： | 李志辉 严律南 赵永恒 刘智敏 刘自明 李玉 |
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| 作者单位： | 四川大学华西医院普外科,成都,610041 |
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| 摘 要： | 目的 构建肝癌特异性HSV—TK／GCV重组腺病毒相关病毒质粒并了解其在细胞内的表达。方法 以腺病毒相关病毒的质粒WAV2作为载体，将TK基因插在甲胎蛋白(AFP)增强子／白蛋白启动子(AFP增强子／ALB启动子)的调控基因的下游，重组成pWAV2／AFP-ALB／HYTK质粒载体。同时构建质粒载体pEGFP-1／AFP—ALB。然后将上述两种不同载体分别转入AFP阳性表达的HepG2细胞株以及阴性表达的7721、SPC和7901细胞株。结果 绿色荧光蛋白只在AFP阳性表达的HepG2细胞株表达，采用PCR技术，以HSV—TK的引物扩增所抽提的总DNA中，只有AFP阳性表达的HepG2细胞株扩增出了710bp的DNA片段。结论 pWAV2／AFP—ALB／HYTK质粒载体在体外实验中具有很好的靶向性。
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| 关 键 词： | 肝癌 特异性HSV-TK／GCV 腺病毒相关病毒 重组质粒 基因治疗 |
| 文章编号： | 1007-9424(2004)03-0234-04 |
| 修稿时间： | 2003-05-27 |
Construction and Study of Specific HSV-TK/GCV Reconstructed Adeno-Associated Virus Plasmid of Human Hepatocellular Carcinoma |
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| Abstract: | Objective To investigate target gene therapy for hepatocellular carcinoma (HCC). Methods Herpes simplex virus thymidine kinase (HSV TK) gene was inserted into the gene of AFP enhancer/ALB promoter with adeno associated virus (AAV) plasmid (WAV2) as a carrier, and a hybrid plasmid pWAV2/AFP ALB/HYTK was constructed. Besides, plasmid pEGFP 1/AFP ALB was also constructed. Two kinds of plasmids were transferred into AFP positive cells HepG2 and AFP negative cells 7721, SPC and 7901.Results It was found that enhance green fluorescence protein could only be seen in AFP positive cells HepG2. 710 bp DNA was amplified only in AFP positive HepG2 cells.Conclusion Plasmid pWAV2/AFP ALB/HYTK for HCC demonstrates specificity in vitro . |
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| Keywords: | Target gene therapy Adeno-associated virus Hepatocellular carcinoma |
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