单链抗体导向卵巢上皮癌进行靶向基因转移的实验研究  

Targeting gene transfer to ovarian epithelial carcinoma cells with lentivirus displaying single-chain antibody.

作　　者：孔北华[1] 宋莉 刘春生[1] 马道新[1] 王瑾瑜[2] 

机构地区：[1]山东医科大学附属医院,250012 [2]北京大学病理生理学教研室

出　　处：《现代妇产科进展》2000年第5期324-327,共4页Progress in Obstetrics and Gynecology

摘　　要：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 targeting gene transfer of Anti MUC 1(ScFv) directed lentivirus to MUC 1 + ovarian epithelial carcinoma cells.Methods:MUC 1 single-chain antibody gene constructed in the plasmid pM.TC.G were transferred to packaging cell line 293T to produce anti MUC 1 directed lentivirus using Ca 3(PO 4) 2 precipitation method. Ovarian epithelial cancer cell line SKOV 3(MUC 1 +) and normal human gingival fibroblast cell GF(MUC 1 -)were co cultured, then were infected by lentivirus. The infection result was observed through fluorescence microscopy. The competition assay that was the addition of the anti MUC 1 single-chain antibody prior to lentivirus infection showed whether anti MUC 1 Ab could inhibite infection. Results: After lentivirus infetion, cell SKOV 3 and GF which were co cultured had remarkably different infection rate. The addition of the MUC 1 Ab prior to the lentivirus infection showed anti MUC 1 Ab can inhibite infection.This competition assay showed that the infection is mediated by the antibody moiety and, therefore, is antibody specific. Conclusions:Anti MUC 1(ScFv) directed lentivirus can targetingly transfer gene into MUC 1 + ovarian cancer cell line.

关 键 词：单链抗体 卵巢肿瘤 细胞系 转染 

分 类 号：R737.31[医药卫生—肿瘤]