单纯疱疹病毒胸苷激酶基因转染联合药物对卵巢癌细胞的杀伤效应

Effect of Herpes Simplex Virus Thymidine Kinase Gene Transduction and Prodrug on Ovarian Cancer Cell

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 evaluate the efficacy of gene therapy with the herpes simplex virus thymidine kinase (HSV TK) gene transference followed by ganciclovir (GCV) therapy in ovarian cancer. Methods: HSV TK gene was inserted into the hind Ⅲ site of pLNSX vector and recombinant plasmid (pLNS/HSV TK) was constracted. Then, the recombinant plasmid was transfered into PA 317 packaged cell and the producer cell line of recombinant retrovirus vector pLNS/HSV TK was established. The cytotoxicity efficacy of HSV TK/GCV system to AO cell line was evaluated by microcucture tetrajolium test (MTT) method. Results: The recombinant retroviral vector pLNS/HSK TK can transfer HSV SK gene into the genome of AO cell line and make it sensitive to GCV. The growth inhibitory rate of AO cell line transfered by pLNS/HSV TK was 98.0% in the medium containing 400μmol GVC. Conclusion: The ovarian cancer cell line transfered by HSV TK gene regressed rapidly in response to the GCV therapy.