基于单克隆抗体G250修饰的肿瘤细胞靶向基因载体研究  

Tumor Targeted Gene Vector Modified with G250 Monoclonal Antibody for Gene Therapy

作　　者：韩素芳[1] 段亚君[1] 王燕铭[1] 郑骏年[2] 武艺[2] 蔡荣[1] 欧来良[1] 孔德领[1] 俞耀庭[1] 

机构地区：[1]生物活性材料教育部重点实验室,南开大学生命科学学院,天津300071 [2]徐州医学院附属医院泌尿外科,徐州221002

出　　处：《高等学校化学学报》2008年第5期923-926,共4页Chemical Journal of Chinese Universities

基　　金：国家自然科学基金(批准号:30370405,20774050);江苏省青年创新人才基金(批准号:BK2005429);卫生部科学研究基金(批准号:WKJ2005-2-026)资助

摘　　要：用宫颈癌细胞Hela表面高表达G250抗原的单克隆抗体G250修饰非病毒基因载体,获得肿瘤靶向基因载体.通过注射G250杂交瘤细胞于小鼠腹腔,制备富含G250mAb的腹水,用正辛酸-硫酸铵沉淀法和Protein A Agarose分离纯化,获得高纯度的G250mAb.通过二硫键将PEI与G250mAb偶联,得到修饰的基因载体G250mAb-PEI,研究其转基因靶向性.结果表明,G250mAb-PEI对Hela细胞的基因转染具有显著的靶向性,对Hela细胞的转基因效率是肝癌细胞HepG2(G250阴性)的2倍;而对正常血管平滑肌细胞(SMC)的基因转染效率比Hela低近20倍,G250mAb修饰与否对SMC没有靶向性;对3T3细胞的毒性显著低于未修饰的PEI,表明G250mAb-PEI是一种高效、低毒和具有靶向性的基因载体.The present study developed clonal antibody. G250mAb can specially a tumor targeted gene vector by modification of PEI with G250 monocombine with the G250 which is a tumor associated with antigen expressed highly in Hela cells. G250mAb was prepared from the ascites of Balb/c mice, and conjugated to PEI via disulfide bonds and generated G250mAb-PEI conjugate. G250mAb-PEI can condense plasmid DNA and form G250mAb-PEI/DNA complex, which can protect DNA from DNaseⅠ digestion. Targeting effect and transfection efficiency of G250mAb-PEI was evaluated via co-culture technology. The results demonstrate that G250mAb-PEI can specially target the Hela cells. The transfection efficiency to Hela is two folds of HepG2 which was G250 negative. The tumor targeting effect was also demonstrated in transfection of smooth muscle cells（SMC）. The transfection efficiency of SMC is almost 20 folds lower than that of Hela cells. In addition, the cytotoxicity of G250mAb-PEI which was determined by MTF assay on NIH 3T3 cells was much lower than PEI. In summary, G250mAb-PEI is a highly efficient gene vector with a low cytotoxicity and targeting effect. More work need to be done to evaluate the potential of the vector in vivo gene therapy.

关 键 词：聚乙烯亚胺 G250mAb 肿瘤靶向 HELA细胞 基因治疗 

分 类 号：Q782[生物学—分子生物学] O623.73[理学—有机化学]