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目的研究腺病毒伴随病毒（ＡＡＶ）载体介导杀肿瘤基因的转移及其在肿瘤治疗方面的应用。方法应用作者构建的腺病毒伴随病毒载体，克隆Ｉ型单纯疱疹病毒胸苷激酶（ＨＳＶＩ－ＴＫ）基因，构建质粒ｐＡＣＴＫ－１９。用ｐＡＣＴＫ－１９转染５型腺病毒（Ａｄ５）感染的重组ＡＡＶ包装细胞系ＡＥ１２０１，获得重组病毒ｒＡＡＶ／ＡＣＴＫ。再用此重组病毒感染肺癌细胞系Ａ５４９，并联合丙氧鸟苷（ＧＣＶ）作用，研究其体外对肺癌细胞的杀伤作用。结果重组ｒＡＡＣ／ＡＣＴＫ的滴度为３４×１０５ＣＦＵ／ｍｌ，感染肺癌细胞系Ａ５４９，实现了ＴＫ基因的转移及与细胞染色体的整合和表达，对Ａ５４９细胞具有杀伤作用。结论腺病毒伴随病毒载体能成功地实施对肺癌细胞系Ａ５４９的基因转移和介导杀肿瘤效应
Objective To study the metastasis of tumor suppressor gene mediated by AAV vector and its application in tumor therapy. Methods Plasmid pACTK-19 was constructed by cloning HSV-TK gene of HSV-I virus. The recombinant AAV packaging cell line AE1201 infected with adenovirus type 5 adenovirus (Ad5) was transfected with pACTK-19 to obtain recombinant virus rAAV / ACTK. The recombinant adenovirus was then used to infect lung cancer cell line A549 in combination with gonadotrophin (GCV) to study its killing effect on lung cancer cells in vitro. Results The titer of recombinant rAAC / ACTK was 3.4 × 10 5 CFU / ml and the lung cancer cell line A549 was infected. The transfer of TK gene and the integration and expression of TK gene with the chromosome of the cell were achieved. Conclusions Adenovirus-associated viral vectors can successfully carry out gene transfer and mediate tumor-killing effect in lung cancer cell line A549