γ-干扰素基因的导入及删除对人肿瘤细胞HLA-Ⅰ、Ⅱ类抗原表达的影响  

The Influence of Transduction and Deletion of IFN-γ over HLA-Class Ⅰ and Ⅱ Antigens, Expression in Human Tumor Cells

作　　者：张腾飞[1] 陈诗书[1] 

机构地区：[1]上海第二医科大学人类基因治疗研究中心,200025

出　　处：《胃肠病学》1999年第1期6-7,共2页Chinese Journal of Gastroenterology

摘　　要：目的：制备有效的细胞因子基因修饰的肿瘤细胞作为“瘤苗”。方法：用缺陷型逆转录病毒作为载体将人γ－干扰素（ＩＦ－γ）含信号肽ｃＤＮＡ转导入人胃癌细胞株ＭＫＮ、４５，经Ｇ４１８筛选后得－ＩＦＮ－γ基因修饰株ＭＫＮ－４５－ＩＦＮ。ＰＣＲ方法测得有标记基因ＮｅｏＲ及ＩＦＮ－γ基因的存在，但经３个月连续传代培养后，ＰＣＲ仅能检测到ＮｅｏＲ基因，而未检出ＩＦＮ－γ基因、ＨＬＡ－Ⅰ、Ⅱ类单抗间接免疫荧光法流式细胞仪测定了上述相应时期ＩＦＮ－γ基因修饰肿瘤细胞ＨＬＡ－Ｉ、Ⅱ类抗原的表达。结果：随着ＩＦＮ－γ基因在染色体中被删除，ＨＬＡ－Ⅰ、Ⅱ类的表达又恢复至与ＭＫＮ－４５相同的水平。结论：为确保ＩＦＮ－γ基因的作用，应不断检测其基因的存在。Background/Aims: To gain the effective cytokine gene modified tumor vaccine. Methods and Results: Retroviral vector pLXSN was used to introduce human cytokine gene IFN-γ into human tumor cell line MKN-45 a modified tumor cell named MKN-45-IFN was gained by G418 screening. PCR was used to detect the IFN gene and mark gene NeoR, but after three months culture, IFN gene could not be found by PCR. Bioanalysis also could not detect the exist of the IFN gene. The IFN gene has been deleted by tumor cell. Conclusions: IFN gene must be detected continuously in order to ensure the reality of expression of HLA-class Ⅰ and Ⅱ antigens.

关 键 词：基因表达 Γ-干扰素基因 HLA-Ⅰ、Ⅱ类抗原 

分 类 号：R730.2[医药卫生—肿瘤]