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用逆转录病毒载体将单纯疱疹病毒胸苷激酶（ＨＳＶｔｋ）基因导人恶性肿瘤细胞，随后应用药物丙氧鸟苷（ＧＣＶ）可选择性地杀死肿瘤细胞．本研究中我们将ＨｙＴＫ基因克隆到逆转录病毒载体ＬＸＳＮ中，并切除其中的ＳＶ４０早期启动子，构建成重组逆转录病毒载体ＬＨｙＴＫ／Ｎ，并用该重组病毒转染小鼠恶性黑色素瘤细胞系Ｂ１６细胞，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，４８ｈ后，在光镜下观察细胞形态及进行活细胞计数．结果表明，ＧＣＶ浓度大于０．１μｍｏｌ／Ｌ即对Ｂ１６／ＨｙＴＫ＋细胞有显著的杀伤作用
Herpes simplex virus thymidine kinase (HSVtk) gene was introduced into malignant cells using retrovirus vectors, and then the drug cytosine (GCV) was used to selectively kill tumor cells. In this study, we cloned the HyTK gene into the retroviral vector LXSN and excised the SV40 early promoter to construct the recombinant retroviral vector LHyTK/N and transfected the mouse malignant melanoma cell line B16 with the recombinant virus. Cells, detected by PCR methods, proved that the HyTK gene has been successfully introduced into tumor cells and does not contain replicable helper virus. HTK and HyTK+ B16 cells were treated with different concentrations of GCV. After 48 hours, the morphology of the cells was observed under light microscope and viable cell counts were observed. The results showed that GCV concentration greater than 0.1 μmol/L had significant killing effect on B16/HyTK+ cells.