联合IL-2基因和B7-1基因转染G422细胞体外生物学特性

Biological characteristics of IL-2 and B7-1 gene co-transfected glioblastoma cells mediated by adenovirus

目的 通过研究联合基因转染后胶质母细胞瘤G42 2细胞生物学特性的变化 ,探讨脑胶质瘤细胞因子基因治疗的新途径。方法 通过重组腺病毒作载体 ,将IL 2基因和B7 1基因转染至小鼠G42 2细胞 ,通过CTLL 2细胞增殖的MTT法检测IL 2的分泌水平 ,FACS检测B7 1和ICAM 1的表达 ,MTT法检测转基因后G42 2细胞的增殖活性 ,双层琼脂糖培养法检测其克隆形成能力。结果 IL 2基因转染 4小时后即可检测到IL 2的分泌 ,2 4小时达高峰 ( 4 83U/ml) ,一周时表达明显下降 ,B7 1高表达 ,ICAM 1表达增加 ,增殖能力和克隆形成能力无变化。结论 重组腺病毒作载体 ,IL 2基因及B7 1基因共转染G42 2细胞 ,能得到目的基因的高效表达 ,同时促进了粘附分子ICAM 1的表达 ,体外增殖活性不受影响。

Objective To seek a new way of gene therapy for brain glioma by studying on IL-2 gene and B7-1 gene cotransfected G422 murine glioblastoma cells.Methods IL-2 gene and B7-1 gene were co-transfected into G422 murine glioblastoma cells mediated by recombinant adenovirus in vitro.The proliferation of G422 cells was tested by MTT method and the level of IL-2 secreted by gene modified G-422 cells was tested by MTT method of the proliferation of CTLL-2.The expression of B7-1 and ICAM-1 on gene transfected G422 cells were assayed by FACS.Their colon-forming ability was tested with bilayer agrose culture after gene transfection.Results 4h after IL-2 gene transfection,IL-2 could be detected in the supernatant of gene modified G422 cells,and the level of IL-2 reached the highest level from 24 to 72 hours,but decreased markedly one week later.B7-1 was highly expressed and the expression of ICAM-1 was enhanced.Conclusion IL-2 gene and B7-1 gene could be co-transfected into G422 cells and could be expressed effectively mediated by recombinat adenovirus,which results in enhanced expression of ICAM-1 of the cells,whereas the proliferation of gene modified G422 cells is not changed.