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目的 探讨抗MUC1单链抗体 (scFv)导向的慢病毒介导的单纯疱疹病毒结构蛋白VP2 2和胸苷激酶 (TK)融合基因及丙氧鸟苷 (GCV)自杀基因系统对MUC1+ 人卵巢上皮癌的特异靶向性生长抑制作用。方法 将慢病毒包装质粒、包膜质粒、转移载体质粒采用磷酸钙沉淀法共转染包装细胞2 93T ,收集病毒上清 ,并建立MUC1+ 人卵巢上皮癌 (3AO)细胞株腹腔移植瘤模型。单药组分为scFv VP2 2 TK +生理盐水 (NS)组、VP2 2 TK +NS组、NS +NS组 ,分别用慢病毒scFv VP2 2 TK、VP2 2 TK或NS1ml腹腔注射 ,继予NS腹腔注射 ;联合用药组分为scFv VP2 2 TK +GCV组、VP2 2 TK +GCV组、NS +GCV组分别应用慢病毒scFv VP2 2 TK、VP2 2 TK及NS腹腔注射 ,2 4h后给予GCV腹腔注射治疗。每组裸小鼠均为 5只。观察各组裸鼠的生存时间及慢病毒的毒性作用。结果 平均生存时间scFv VP2 2 TK +NS组、VP2 2 TK +NS组、NS +NS组、NS +GCV组、VP2 2 TK +GCV组、scFv VP2 2 TK +GCV组分别为 18.4d± 2 .9d、18.8d± 1.5d ,17.6d± 1.1d ,18.5d± 1.6d ,2 4d± 5d和 4 6d± 2 2d ,6组比较 ,差异有显著意义 (χ2 =2 4 .82 ,P =0 .0 0 2 ) ;并且scFv VP2 2 TK +GCV组较VP2 2 TK +GCV组裸鼠平均生存时间明显延长 (χ2 =7.4 3,P =0 .0 0 6 )。结
Objective To investigate the effect of anti-MUC1 scFv-directed lentivirus-mediated herpes simplex virus structural protein VP2 2 and thymidine kinase (TK) fusion gene and the guanosine monophosphate (GCV) suicide gene system on MUC1 + human epithelial ovarian cancer Specific targeted growth inhibition. Methods Lentiviral packaging plasmid, envelope plasmid and transfer vector were cotransfected into 293T cells with calcium phosphate precipitation, and the virus supernatant was collected. The model of peritoneal xenograft in MUC1 + human ovarian epithelial carcinoma (3AO) cell line was established. The single drug components were scFv VP2 2 TK + NS group, VP2 2 TK + NS group and NS + NS group, which were injected intraperitoneally with lentivirus scFv VP2 2 TK, VP2 2 TK or NS1 ml respectively, followed by intraperitoneal injection of NS The VP2 2 TK + GCV group and NS + GCV group were injected with VP2 2 TK, VP2 2 TK and NS of lentivirus respectively. After 24 h, mice were injected intraperitoneally with GCV treatment. Each group of nude mice were 5. The survival time and toxicity of lentivirus in each group were observed. Results The average survival time was 18.4d ± 2 in scFv VP2 2 TK + NS group, VP2 2 TK + NS group, NS + NS group, NS + GCV group, VP2 2 TK + GCV group and scFv VP2 2 TK + GCV group. 9d, 18.8d ± 1.5d, 17.6d ± 1.1d, 18.5d ± 1.6d, 24d ± 5d and 46d ± 2d, there was significant difference between the 6 groups (χ2 = 2.42, P = 0 .0 0 2); and the average survival time of scFv VP2 2 TK + GCV group was significantly longer than that of VP2 2 TK + GCV group (χ2 = 7.43, P = 0.060). Knot