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目的 :探讨靶向性基因转移载体—含抗MUC 1单链抗体的慢病毒对MUC 1+卵巢上皮癌细胞系SKOV 3细胞进行基因转移的特异性 ,为卵巢上皮癌的靶向性基因治疗提供实验基础。方法 :将针对SKOV 3细胞上抗原MUC 1的单链抗体基因构建到慢病毒包膜结构质粒 (pM .TC .G) ,以报告基因绿色荧光蛋白 (GFP)基因作为目的基因。用磷酸钙沉淀法进行Anti MUC 1(ScFv)慢病毒包装 ,感染共培养的卵巢癌细胞SKOV 3(MUC 1+)和正常人成纤维细胞GF(MUC 1- ) ,荧光显微镜下观察感染效果。竞争抑制实验即病毒感染前加入Anti MUC 1单抗是否抑制病毒感染。结果 :荧光显微镜下观察到Anti MUC 1(ScFv)介导的慢病毒对共培养的SKOV 3(MUC 1+)和GF(MUC 1- )细胞的感染有明显差别 ,非Anti MUC 1(ScFv)介导的慢病毒对共培养的SKOV 3(MUC 1+)和GF(MUC 1- )的感染未见显著性差别 ,证明Anti MUC 1(ScFv)介导的慢病毒可对MUC 1+的卵巢细胞靶向性转染。病毒感染前加入Anti MUC 1单抗则可抑制病毒靶向性感染。此竟争抑制实验表明 ,病毒感染是由其抗体组成部分介导 ,故有抗体靶向性。结论 :Ani MUC 1(ScFv)介导的慢病毒可对MUC - 1+卵巢上皮癌细胞靶向性基因转移。
OBJECTIVE: To investigate the specificity of targeted gene transfer vector-lentivirus containing anti-MUC 1 single chain antibody for gene transfer in MUC 1+ epithelial ovarian cancer cell line SKOV 3 and to provide targeted gene therapy for epithelial ovarian cancer Experimental basis. Methods: The single chain Fv gene against MUC 1 of SKOV 3 cells was constructed into the lentivirus envelope plasmid (pM .TC .G) and the reporter gene GFP gene was used as the target gene. The anti-MUC 1 (ScFv) lentivirus was packaged by calcium phosphate precipitation, and the co-cultured ovarian cancer cells SKOV 3 (MUC 1+) and normal human fibroblasts GF (MUC 1-) were infected and the infection effect was observed under a fluorescence microscope. Competition inhibition test means whether Anti MUC 1 McAb is added before virus infection to inhibit virus infection. Results: Anti-MUC 1 (ScFv) -mediated lentivirus showed significant difference in co-cultured SKOV 3 (MUC 1+) and GF (MUC 1-) cells under fluorescence microscope. Mediated lentiviruses showed no significant difference in the co-cultured SKOV 3 (MUC 1+) and GF (MUC 1-) infections, demonstrating that the anti-MUC 1 (ScFv) -mediated lentivirus can inhibit ovarian MUC 1+ Cell-targeted transfection. AntiMUC 1 mAb was added before virus infection to inhibit viral targeted infection. This competitive inhibition test shows that viral infection is mediated by its antibody components and therefore has antibody targeting. Conclusion: Ani MUC 1 (ScFv) - mediated lentivirus can target gene transfer to MUC - 1 + epithelial ovarian cancer cells.