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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 investigate the biological characteristics of G42 2 cells after the combination gene transfection, and to explore new ways of gene therapy for glioma cytokines. Methods IL2 gene and B7 1 gene were transfected into mouse G42 2 cells by recombinant adenovirus vector. IL 2 secretion was detected by MTT assay of CTLL 2 cell proliferation. Expression of B7 1 and ICAM 1 was detected by FACS. The proliferation activity of G42 2 cells was detected by MTT assay and the colony formation ability was detected by double-layer agarose culture. Results The secretion of IL 2 was detected 4 hours after transfection of IL 2 gene, reached the peak at 24 hours (4 83 U / ml), decreased significantly at 1 week, increased at B7 1, increased ICAM 1 expression, proliferation and clonality No change in ability to form. CONCLUSION: Recombinant adenovirus vector can co-transfect IL-2 gene and B7 1 gene into G42 2 cells. The recombinant adenovirus can efficiently express the target gene and enhance the expression of ICAM-1. The proliferative activity in vitro is not affected.