摘要目的 采用大鼠诱发性卵巢癌模型,观察经肿瘤动脉灌注结合GE7导入系统对Ⅰ型单纯疱疹病毒胸腺嘧啶核苷激酶基因(HSV1-tk)转导的效率和靶向性.方法 构建GE7四元复合物,将诱发的荷瘤大鼠9只随机分成A组、B组和C组,A组经卵巢动脉注入四元复合物(8μg/只),B组注入同体积生理盐水,C组经尾静脉注入四元复合物(8μg/只).给药72 h后,分别以逆转录聚合酶链反应(RT-PCR)和Western blot检测大鼠的肿瘤、心、肝、脾、肺及肾脏组织的HSV1-tk mRNA及蛋白的表达.结果 A组肿瘤组织中HSV1-tk mRNA和蛋白均高表达,而其他组织中则无表达或表达极少;B组未见HSV1-tk mRNA及蛋白的表达;C组肿瘤组织中未见HSV1-tkmRNA和蛋白的表达,肝脏、脾脏、肺脏及肾脏组织中则有少量的表达.RT-PCR半定量检测结果显示,A组卵巢肿瘤组织中的HSV1-tk mRNA含量(1.692±0.221)明显高于C组卵巢肿瘤组织(0.012±0.002;P<0.01).结论 动脉灌注结合GE7导入系统对HSV1-tk有较高的转导率和靶向性,为HSV1-tk/GCV治疗系统在卵巢癌基因治疗的应用提供了新的策略.

abstractsObjective To observe the gene and protein expression of herpes simplex virus type Ⅰ thymidine kinase (HSV1-tk) in the ovarian tumor tissues and other organs after arterial infusion of HSV1-tk gene mediated by GE7 delivery system.Methods GE7-polylysine/pCMV-HSV1-tk/polylysine-HA20complexes were constructed.Nine rats with induced ovarian tumor were divided into 3 groups, injecting the 4-element complexes or saline buffer through the ovarian artery and complexes through the tail vein,respectively.The ovarian tumors, hearts, livers, spleens, lungs and kidneys were obtained at 72 hours after injection.RT-PCR and Western Blot were preceeded to determine the expression of HSV1-tk gene and protein in the tumor tissues and other organs.Results In the group of arterial injection with 4-element complexes, the HSV1-tk gene and protein were expressed strongly in the tumor tissues, while little or none was detected in other organs.In the group of arterial injection with saline buffer, no HSV1-tk gene and protein was detected in both tumor tissues and other organs.In the group of tail vein injection, none was detected in tumor tissues and only little was found in the livers, spleens, lungs and kidneys.Conclusion High target and gene transfer rates can be obtained when HSV1-tk gene is transferred via the artery route mediated by GE7 delivery system.HSV1-tk protein can be expressed after the gene transfer.The results may provide a new strategy for target killing of HSV1-tk/GCV system in ovarian tumors.