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目的：探讨单纯疱疹病毒胸苷激酶（ＨＳＶ－ＴＫ）基因转染联合抗病毒药物羟基无环鸟苷（ＧＣＶ）对人卵巢癌细胞系的杀伤效应。方法：采用基因工程技术，将ＨＳＶ－ＴＫ基因的ＤＮＡ序列插入逆转录病毒载体ｐＬＮＳＸ的ＨｉｎｄⅢ位点，构建重组体ｐＬＮＳ／ＨＳＶ－ＴＫ，并采用磷酸钙介导贴壁细胞基因转染法将重组体导入ＰＡ３１７包装细胞，建立重组逆转录病毒载体分泌细胞株ＰＡ３１７／ＴＫ。并采用四甲基偶氮唑蓝比色法检测ＨＳＶ－ＴＫ／ＧＣＶ系统对人类卵巢癌ＡＯ细胞系的生长抑制率。结果：重组逆转录病毒载体ｐＬＮＳ／ＨＳＶ－ＴＫ能有效地将ＨＳＶ－ＴＫ基因导入ＡＯ细胞系内并使其获得对ＧＣＶ的敏感性。当培养液内ＧＣＶ达４０μｍｏｌ时，其对转ＨＳＶ－ＴＫ基因后ＡＯ细胞的生长抑制率为９８．０％。结论：ＡＯ细胞转染ＨＳＶ－ＴＫ基因后，能有效地被ＧＣＶ杀灭。
Objective: To investigate the killing effect of herpes simplex virus thymidine kinase (HSV-TK) gene combined with antiviral drug acyclovir (GCV) on human ovarian cancer cell lines. METHODS: The DNA sequence of HSV-TK gene was inserted into the HindIII site of retroviral vector pLNSX by using genetic engineering to construct recombinant pLNS / HSV-TK. Recombinant pLNS / HSV-TK was constructed by using calcium phosphate-mediated adherent cell transfection PA317 packaging cells into the recombinant retroviral vector-secreting cell line PA317 / TK. The growth inhibition rate of HSV-TK / GCV system on human ovarian cancer AO cell line was detected by MTT assay. Results: The recombinant retroviral vector pLNS / HSV-TK effectively introduced HSV-TK gene into AO cell line and made it sensitive to GCV. When culture medium GCV up to 40μmol, its inhibition of HSV-TK transfected AO cells growth inhibition rate of 98.0%. Conclusion: AO cells can be effectively killed by GCV after transfection with HSV-TK gene.