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用逆转录病毒载体将单纯疱疹病毒胸苷激酶基因（ＨＳＶｔｋ）导入恶性肿瘤细胞，随后可应用药物９－（１，３－二羟基－丙氧基－甲基）鸟嘌呤（ｇａｎｃｉｃｌｏｖｉｒ，ＧＣＶ）选择性地杀死肿瘤细胞．将ＨｙＴＫ基因替换逆转录病毒载体ＧｌＮａ中的ｎｅｏ基因，构建成重组逆转录病毒载体ＧＴＫ，转染混合包装细胞（双噬性ＰＡ３１７细胞和单噬性ＧＰ＋Ｅ－８６细胞），通过“乒乓效应”获得高滴度重组病毒．用该重组病毒转染小鼠恶性黑色素瘤细胞系Ｂ１６细胞，用ｈｙｇｒｏｍｙｃｉｎＢ筛选出阳性细胞克隆（ＨｙＴＫ＋），经ＰＣＲ方法检测证明ＨｙＴＫ基因已成功地导入肿瘤细胞中，且不含可复制的辅助病毒．分别用不同浓度的ＧＣＶ作用于ＨｙＴＫ－及ＨｙＴＫ＋的Ｂ１６细胞，光镜下观察２４ｈ和４８ｈ后细胞形态及进行活细胞计数．结果表明，ＧＣＶ浓度大于０．１μｍｏｌ／Ｌ时即对Ｂ１６／ＨｙＴＫ＋细胞有显著的杀伤作用
Herpes simplex virus thymidine kinase gene (HSVtk) is introduced into malignant cells using a retroviral vector, followed by selection of the drug ga-ciclovir (GCV) Kill tumor cells sexually. The HyTK gene was substituted for the neo gene in the retroviral vector GlNa into a recombinant retroviral vector GTK and transfected into mixed packaging cells (Biphasic PA317 cells and monocytic GP + E-86 cells) by “ping-pong effect” High titers of recombinant virus. The murine malignant melanoma cell line B16 was transfected with the recombinant virus, and the positive cell clone (HyTK +) was screened with hygromycin B. The HyTK gene was proved to be successfully introduced into tumor cells by PCR and contained no replicable helper virus . Cells were treated with different concentrations of GCV in HyTK- and HyTK + B16 cells respectively. The cell morphology was observed 24h and 48h under light microscope and the viable cells were counted. The results showed that GCV concentration greater than 0.1μmol / L when B16 / HyTK + cells have a significant killing effect