逆转录病毒介导人白细胞介素2基因在人膀胱癌细胞中高效表达

Retrovirus-mediated human IL-2 gene transfer in human bladder carcinoma cells

构建人白细胞介素２（ｈＩＬ－２）基因逆转录病毒表达载体ｐＺｉｐ－ｈＩＬ－２，用ＰＡ３１７包装细胞系包装后获得高效价逆转录病毒（１～５×１０５ｃｆｕ／ｍｌ）。病毒感染人膀胱癌细胞系ＢＩＵ８７并用Ｇ４１８选择培养，得到Ｇ４１８抗性克隆；经小鼠脾母细胞增殖法测定、ＲＮＡＳｌｏｔ打点杂交实验和Ｗｅｓｔｅｒｎ免疫印迹实验证实其高效表达ｈＩＬ－２，平均活性在２４小时每毫升为７５Ｕ／１０６细胞（最高达１２５Ｕ／ｍｌ）。转ｈＩＬ－２基因在人膀胱癌细胞中的高效表达，可为人膀胱癌转基因瘤苗的应用研究提供实验基础。

Retrovirus-mediated human IL-2 gene expression vector pZip-hIL-2 has beeri developed.After packaged by PA317 packaging cell line,the retrovirus was obtained at a high titer(1～5×10_5cfu/ml),and infected the human bladder cancer line BIU87.On G418-selecting culture, G418-resistant colonies were obtained and hIL-2 was highly expressed up to 125U/ml as detected by IL-2 activity assay,western blotting and RNA Slot hybridization.This laid the foundation to develop IL-2 gene-transduced tumor vaccine of human bladder cancer.