转人GM—CSF基因人膀胱癌细胞株的建立及体外生物学特性

经逆转录病毒载体将人GM-CSF基因导入人膀胱癌细胞株BIU-87细胞中,建立了转基因细胞株BIU/GM。经流式细胞仪行细胞DNA周期分析表明GM-CSF基因的导人及表达对BIU-87细胞的增长无影响。免疫荧光测定发现转GM-CSF基因及表达不能促进BIU-87细胞表面HLA-ABC、DR、DQ抗原的表达。转基因瘤细胞株经6000rad X射线照射灭活后,丧失增殖能力,逐步死亡,但能维持一定水平的GM-CSF分泌活性达两周以上。从而为制备灭活的转基因瘤苗提供了初步经验。

Establishment of Human Bladder Cancer Cell Line Transfected with hGM-CSF Gene and Its biological Characteristics in vitro

Presented in this paper was the report of possibility of bladder cancer cell vaccine transfected with hGM-CSF. The hGM-CSF gene was transferred to the human bladder cancer line cell line B1U-87 by retroviral vector N2A/CMV/hGM-CSF. After screening by G418 and detecting secretory activity, transfected gene cell line BIU/GM wis established. PCR and Southern blot demonstrated that. GM-CSF gene had been integrated into the BIU/GM cell genome. The cell DNA analysis by flowcytometty indicated that, the expression of GM-CSF gene had no influence on the growth and proliferation of BIU cells. Fluorescent im-munoassay revealed that transfected GM-CSF gene and its expression could not promoted the expression of HLA-ABC, DR and DQ antigens on the surface of the gene transfected BIU-87 cells. After the gene transfected BIU cells were irradiated with 6000 rad X-ray, they could secrete hGM-CSF for at least two weeks, although the growth of them wat abrogated. These suggested that we coutd develop the tumor vaccine secreting hGM-CSF for clinical trials in future.