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[目的]通过腺病毒转染的方法将hGM-CSF基因转染到人纤维肉瘤细胞株中制成瘤细胞疫苗。[方法]用携带hGM-CSF基因的腺病毒转染人纤维肉瘤细胞株,并用RT-PCR和ELISA的方法检测目的基因在mRNA水平和蛋白水平的表达。再用射线照射经转染基因实验细胞株从而失活制成肿瘤疫苗。[结果]携带hGM-CSF基因的腺病毒能够成功转染纤维肉瘤细胞,转染的目的基因在mRNA水平的表达在转染后48h达到最高,在蛋白水平的表达可以持续3周以上。[结论]以腺病毒为载体的基因转染系统可以成功建立能分泌hGM-CSF细胞因子的人纤维肉瘤瘤细胞疫苗。
[Objective] The purpose of this study was to transfect hGM-CSF gene into human fibrosarcoma cell line by adenovirus transfection to make tumor cell vaccine. [Methods] Human fibrosarcoma cell lines were transfected with adenovirus carrying hGM-CSF gene. The expression of the target gene at mRNA and protein level was detected by RT-PCR and ELISA. And then irradiated with the transfected cells transfected cells to inactivate the tumor vaccine made. [Result] The adenovirus carrying hGM-CSF gene could successfully transfect fibrosarcoma cells. The expression of the transfected target gene at the mRNA level reached the highest at 48h after transfection, and the expression at the protein level could last for more than 3 weeks. [Conclusion] Adenovirus-mediated gene delivery system can successfully establish human fibrosarcoma cell vaccine capable of secreting hGM-CSF cytokines.