转hGM－CSF基因人膀胱癌细胞株的建立及其生物学特性

Estabolishment of Cell Line of Human Bladder Cancer Transfected with hGM CSF Gene and its Biological Characteristics

采用逆转录病毒载体将ｈＧＭ－ＣＳＦ基因导入人膀胱癌细胞株ＢＩＵ－８７细胞中，建立转基因细胞株ＢＩＵ／ＧＭ。经ＰＣＲ－ＳｏｕｔｈｅｒｎＢｌｏｔ杂交证实ＧＭ－ＣＳＦ基因已整合到ＢＩＵ／ＧＭ细胞基因组中。经流式细胞仪细胞ＤＮＡ周期分析表明，转基因前后ＢＩＵ－８７细胞增殖状态无变化。免疫荧光检测发现ＧＭ－ＣＳＦ基因导入及表达不能促进ＢＩＵ－８７细胞表面ＨＬＡ－ＡＢＣ、ＤＲ、ＤＱ抗原的表达。转基因细胞经６０Ｇｙ／ｓＸ线灭活后，丧失增殖能力，但能维持一定水平的ＧＭ－ＣＳＦ分泌活性达２周。本文为进行转基因瘤苗的深入研究奠定了初步基础。

The cDNA of hGM CSF was transfered into the human bladder cancer line BIU 87 with retroviral vector. PCR southern blot demonstrated the integration of transfered gene in the BIU cell genome. Biological activity of the hGM CSF secreted by the BIU cell was confirmed. DNA analysis by flowcytomerty showed no influence on growth of the BIU cells. Flourenscent immunoassay revealed no induction of HLA ABC, DR, DQ expression on the surface of the gene transfered BIU cells. The gene transfected BIU cells were irradiated with 6000 rad X ray but they could secret hGM CSF for at least 2 weeks, although the growth of them was abrogated. These suggested that we could develope the bladder cancer vaccine secreting hGM CSF for clinical trials in future.