CD71抗体靶向转移系统的构建及对细胞的生长抑制作用  

THE CONSTRUCTION OF A TARGETING DELIVERY SYSTEM MEDIATED BY CD71 ANTIBODY AND IT'S INHIBITION EFFECT ON THE GROWTH OF HEPG2 CELLS

作　　者：龚道元[1] 郭婷婷[2] 佟豪 伏红霞[2] 彭艳[2] 王晓娟[1] 

机构地区：[1]广东省佛山科学技术学院医学院检验系,佛山528000 [2]内蒙古医学院免疫学教研室,呼和浩特010059 [3]内蒙古肿瘤医院肿瘤内科,包头014000

出　　处：《广西医科大学学报》2010年第5期684-686,共3页Journal of Guangxi Medical University

基　　金：教育部"春晖计划"资助项目(No.Z2005-2-01007)

摘　　要：目的:构建肝癌靶向性的Survivin反义RNA/HSP70基因转移系统,并研究其在体外对肝癌细胞的生长抑制作用。方法:体外培养抗转铁蛋白受体(CD71)抗体的杂交瘤细胞株D2C5.6,诱导裸鼠产生腹水并纯化,以水溶性碳二亚胺介导CD71抗体与多聚左旋赖氨酸(PLL)的连接,再与Survivin反义RNA/HSP70双基因载体按比例混匀,得到-PLL-载体系统;转染人肝癌细胞株HepG2后,采用MTT法观察细胞的生长情况。结果:分析型酸性尿素凝胶电泳实验证实抗CD71抗体与PLL成功连接;MTT试验显示,Ab-PLL-Survivin反义RNA/HSP70基因转移系统处理的HepG2细胞生长明显受到抑制。结论:CD71抗体介导的肝癌靶向转移系统已经成功构建,为肝癌的基因治疗提供了一定的实验基础。Objective:To construct the gene targeting delivery system mediated by CD71 antibody,and to investigate it's inhibition effect on human liver cancer cell line HepG2.Methods:We injected a number of murine hybridomas D2C5.6 into abdominal cavities of nude mouses and got enough ascitic fluid and purified it.Then it was coupled to poly-L-lysine mediated by EDC.The double-gene expression vector(pIRES2-Survivin antisense RNA/HSP70) was complexed to the conjugate after determine the combination ratio of plasmid and Ab-PLL to form a complex including three components.MTT method was used to measure cell growth after transfection.Results:CD71 antibody was coupled to PLL by analyzing acidic carbamide eletrophoresis.HepG2 cells treated by Ab-PLL-Survivin antisense RNA/HSP70 gene delivery system was inhibited significantly.Conclusion:A targeting delivery system mediated by CD71 antibody was constructed successfully,and it provided experimental basis for gene therapy of liver cancer.

关 键 词：CD71 抗体靶向 基因转移系统 构建 人肝癌细胞株 生长抑制作用 CELLS THE GROWTH Survivin 抗转铁蛋白受体 多聚左旋赖氨酸 HEPG2细胞生长 杂交瘤细胞株 PLL 双基因载体 肝癌靶向性 载体系统 系统处理 体外培养 碳二亚胺 

分 类 号：R[医药卫生]