逆转录病毒介导人白细胞介素2基因在人膀胱癌细胞中的表达

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

用ＰＡ３１７包装细胞系包装人白细胞介素２（ｈＩＬ－２）基因逆转录病毒表达载体ｐＺｉｐ－ｈＩＬ－２后，获得高效价逆转录病毒（０．４～１．２）×１０５ｃｆｕ／ｍｌ。病毒感染人膀胱细胞系ＥＪ，通过Ｇ４１８抗性筛选及活性检测，得到一株高分泌ｈＩＬ－２（２６０Ｕ／ｍｌ）克隆ＥＪ／ｈＩＬ－２，经Ｓｏｕｔｈｅｒｎ印迹分析证实ｈＩＬ－２基因已整合到ＥＪ／ｈＩＬ－２基因组中。转ｈＩＬ－２基因在人膀胱癌细胞中的高效表达，为人膀胱癌转ｈＩＬ－２基因瘤苗的应用研究打下了基础。

After retrovirus mediated human IL 2 gene expression vector pZip hIL 2 was packaged by PA 317 pcaking cell line, the retrovirus was obtained at a high titer (0.4-1.2×10 5 cfu/ml),and infected the human bladder cancer line EJ, On G418 resistance selection and hIL 2 activity assay, a clone (named EJ/hIL 2) secreting highest level of hIL 2(260 U/ml) was selected ,and then Southern blot hybridization demonstrated that human IL 2 gene was integrated in the genome of EJ/hIL 2 cells. This laid the foundation to develop hIL 2 gene transducted tumor vaccine of human bladder cancer.