逆转录病毒载体介导人GM－CSF基因转染肿瘤细胞

应用逆转录病毒载体ｐＺＩＰＮｅｏＳＶ（Ｘ）介导人ＧＭ－ＣＳＦ基因转染肿瘤细胞获得表达。经Ｌｉｐｏｆｅｃｔｉｎ将含有人ＧＭ－ＣＳＦ基因的重组逆转录病毒载体ｐＺＩＰ－ＧＭ转染兼性病毒包装细胞系ＰＡ３１７，继之用病毒收获液感染人肝癌细胞ＳＭＭＣ７７２１和人胃癌ＢＧＣ－８２３，经ＧＭ－ＣＳＦ依赖细胞株ＴＦ１测活和双抗体夹心法ＥＬＩＳＡ测定表明：人ＧＭ－ＣＳＦ基因在人肿瘤细胞中获得稳定高效表达。为进─步建立ＧＭ－ＣＳＦ的转基因治疗模型提供了基础。

RETROVIRAL-MEDIATED TRANSFER OF THE HUMAN GMCSF GENE INTO TUMOR CELLS

Expression of GM-CSF gene in the tumor cells has been obtained with the retroviral vector-mediated gene tranfer method. Used zip-GM, a recombinant retroviral vector containing the GM-CSF gene,to transfect amphotropic and Ecotropic virus packing cell line by lipofectin.Subsequently.cell-free culture supernatant was used to infect human gastric carcinoma cell BGC-823 and human liver tumor cell SMMC7721. Biology activity assay with GM-CSF.dependant cell line TF-1 and two-antibody sandwich ELISA showed that the human GM-CSF gene can stably and highly express in human tumor cells. Thus,this study lays the foundation for further studying tumor gene therapy of GM-CSF.