目的：建立基因枪介导的基因转移系统，为基因修饰的肿瘤细胞疫苗研究奠定实验基础。方法：采用基因枪转导的方法，将真核表达质粒（pcDNA3.1GM-CSF）转导入人胃癌细胞株（SGC），经G418筛选获得阳性克隆(SGC-GM-CSF)，通过RT-PCR方法鉴定，重组载体已整合到胃癌细胞的基因组中。结果： SDS-PAGE和Western blot分析的SGC-GM-CSF上清液结果显示rhGM-CSF主带在30 kD左右，SGC-GM-CSF在体外长期培养中保持稳定分泌，分泌量达到24 h平均247 ng/10 6 cell。结论：建立的基因枪介导的基因转移系统安全、有效，为应用于肿瘤基因治疗提供了实验基础。

Objective:To provide an effective hGM-CSF gene transferring vector mediated by gene gun and a basis for study of hGM-CSF gene-modified tumor cell vaccines. Method: The gastric tumor cell line (SGC) was transfected with eukarytic expression plasmid deoxyribonucleic acid containing the human granulocyte-macrophage colony-stimulating (hGM-CSF) gene using the gene gun. The SGC cell clones (SGC-GM-CSF-1～5)secreting high hGM-CSF level were obtained after G418 resistance selection. The hGM-CSF gene had been integrated into chromatosome of SGC by the assay of RT- PCR.Results: There was hGM-CSF production whose lane was about 30 kD in the culture medium of SGC-GM-CSF by the assay of SDS-PAGE and Western blot. SGC-GM-CSF had the ability of the high level of GM-CSF for a long-time(mean 247ng/(10 6 cell·24 h).Conclusion: The hGM-CSF gene transferring vector mediated by gene gun was effective and safe. These results provide a basis for study of GM-CSF gene therapy for cancer.

本课题是福建省自然科学基金项目(C97067)资助