基因疫苗pcDNA3.0-PSMA肿瘤细胞模型的建立  

Establishment of the Tumor Cells Model with pcDNA3.0-PSMA for DNA vaccine

作　　者：汪波[1] 曹开源[2] 徐向东[3] 徐霖[4] 袁广卿[2] 张甜[2] 丘少鹏[5] 

机构地区：[1]中山大学附属第一医院检验医学部,广州510080 [2]中山大学中山医学院临床检验标准化研究中心,广州510080 [3]中山大学附属第一医院血管外科,广州510080 [4]中山大学中山医学院免疫学教研室,广州510080 [5]中山大学附属第一医院泌尿外科,广州510080

出　　处：《热带医学杂志》2008年第7期639-641,656,F0003,共5页Journal of Tropical Medicine

基　　金：国家自然科学基金(No.30772503;No.30371426);广东省自然科学基金(No.021907)

摘　　要：目的构建表达前列腺特异性膜抗原(PSMA)的肿瘤细胞模型,为基因疫苗的抑瘤效应研究及免疫机制的探讨提供实验材料。方法脂质体转染法分别将PSMA-pcDNA3.0质粒和pcDNA3.0质粒转染至SP2/0细胞,G418筛选后获得了稳定生长的阳性克隆细胞株,RT-PCR、间接免疫荧光法、Westernblot检测PSMA蛋白的表达。结果RT-PCR、间接免疫荧光法和Western blot均证明转染了PSMA-pcDNA3.0质粒的SP2/0细胞表达PSMA。结论成功建立了稳定表达PSMA的小鼠肿瘤细胞模型,最终为前列腺癌的预防和免疫治疗研究打下了坚实的基础。Objective To construct the tumor cells model expressing prostate-specific membrane antigen （PSMA） for the research of anti-tumor effect and immune mechanism about the DNA vaccine. Methods Small-scale preparation of purified pcDNA3.0 and pcDNA3.0-PSMA plasmids were stably transfected into SP2/0 cell lines using Lipofectamine 2000TM. These transfected cells were cultured by G418 （500 μg/mL）. Those survival cells were detected for the expression of PSMA by the RT-PCR, indirect immunofluorescence and Western blot. Results The results of RT-PCR, indirect immunofluorescence and Western blot all demonstrated the SP2/0-PSMA cells had been obtained and could express PSMA. Conclusion The successful construction of the SP2/0-PSMA cells provided the materials for the research of the anti-tumor efficacy and the immune mechanism.

关 键 词：前列腺特异性膜抗原 基因疫苗 基因表达 

分 类 号：R737.25[医药卫生—肿瘤]